HPLC methods for the analysis of fat-soluble vitamins in feeds
\\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:"3447",leadTitle:null,fullTitle:"Psoriasis - Types, Causes and Medication",title:"Psoriasis",subtitle:"Types, Causes and Medication",reviewType:"peer-reviewed",abstract:"By virtue of the dynamic nature of the scientific process, the description of the type, pathogenesis and treatment of a disease is always a work in progress. 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\r\n\tWearable technology and disruptive innovation have transformed the way to look at the devices used in day-to-day life. It is one of the most important interdisciplinary domains which is being used promptly in healthcare, textile, electronics, and defense areas. In healthcare, the focus is more on developing cost-effective point of care wearable devices that can further be integrated with a smartphone for wireless data storage and analysis. The same approach has also been used in textile to develop intelligent garments. The development of new material systems, designs, and fabrication facilities have further strengthened this technology.
\r\n\r\n\tThe book will provide a deep insight into wearable technology for smart textile, defense, healthcare, and fitness applications, including wearable electronics circuits and devices. It will cover the latest research works carried out in materials, design, and fabrication technologies. Thus, the book “Wearable Technology” will be an exclusive reference material for academicians, researchers, and industry personnel (healthcare, textile, and electronics) working towards advancement in this inter-disciplinary area.
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He is the recipient of the prestigious Quick Hire Fellowship and Senior Research Fellowship from CSIR. He is a member of OSA (Optical Society of America), and IAENG (International Association of Engineers) organizations. He is also an author and reviewer for many journals, working as a Postdoctoral Fellow at The Innovative Technologies Laboratory, Saudi Arabia.",coeditorOneBiosketch:"Dr. Kumar received his Ph.D. from Delhi Technological University where he was awarded commendable research excellent awards. He is currently an Assistant Professor in the Electronics and Communication Engineering Department at Jaypee Institute of Information Technology, a Junior Research Fellow at Delhi Technological University, and a visiting faculty in the Department of Electronics and Communication at the Netaji Subhas University of Technology. 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Currently, he is working as an Postdoctoral Fellow at The Innovative Technologies Laboratory (ITL) in KAUST, Saudi Arabia.",institutionString:"The Innovative Technologies Laboratory (ITL) in KAUST",position:null,outsideEditionCount:0,totalCites:0,totalAuthoredChapters:"0",totalChapterViews:"0",totalEditedBooks:"0",institution:null}],coeditorOne:{id:"464519",title:"Dr.",name:"Ajay",middleName:null,surname:"Kumar",slug:"ajay-kumar",fullName:"Ajay Kumar",profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:"Jaypee Institute of Information Technology",position:null,outsideEditionCount:0,totalCites:0,totalAuthoredChapters:"0",totalChapterViews:"0",totalEditedBooks:"0",institution:{name:"Jaypee Institute of Information Technology",institutionURL:null,country:{name:"India"}}},coeditorTwo:null,coeditorThree:null,coeditorFour:null,coeditorFive:null,topics:[{id:"11",title:"Engineering",slug:"engineering"}],chapters:null,productType:{id:"1",title:"Edited Volume",chapterContentType:"chapter",authoredCaption:"Edited by"},personalPublishingAssistant:{id:"347259",firstName:"Karmen",lastName:"Daleta",middleName:null,title:"Ms.",imageUrl:"//cdnintech.com/web/frontend/www/assets/author.svg",email:"karmen@intechopen.com",biography:null}},relatedBooks:[{type:"book",id:"10198",title:"Response Surface Methodology in Engineering Science",subtitle:null,isOpenForSubmission:!1,hash:"1942bec30d40572f519327ca7a6d7aae",slug:"response-surface-methodology-in-engineering-science",bookSignature:"Palanikumar Kayaroganam",coverURL:"https://cdn.intechopen.com/books/images_new/10198.jpg",editedByType:"Edited by",editors:[{id:"321730",title:"Prof.",name:"Palanikumar",surname:"Kayaroganam",slug:"palanikumar-kayaroganam",fullName:"Palanikumar Kayaroganam"}],productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"1591",title:"Infrared Spectroscopy",subtitle:"Materials Science, Engineering and Technology",isOpenForSubmission:!1,hash:"99b4b7b71a8caeb693ed762b40b017f4",slug:"infrared-spectroscopy-materials-science-engineering-and-technology",bookSignature:"Theophile Theophanides",coverURL:"https://cdn.intechopen.com/books/images_new/1591.jpg",editedByType:"Edited by",editors:[{id:"37194",title:"Dr.",name:"Theophile",surname:"Theophanides",slug:"theophile-theophanides",fullName:"Theophile Theophanides"}],productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"3161",title:"Frontiers in Guided Wave Optics and Optoelectronics",subtitle:null,isOpenForSubmission:!1,hash:"deb44e9c99f82bbce1083abea743146c",slug:"frontiers-in-guided-wave-optics-and-optoelectronics",bookSignature:"Bishnu Pal",coverURL:"https://cdn.intechopen.com/books/images_new/3161.jpg",editedByType:"Edited by",editors:[{id:"4782",title:"Prof.",name:"Bishnu",surname:"Pal",slug:"bishnu-pal",fullName:"Bishnu Pal"}],productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"371",title:"Abiotic Stress in Plants",subtitle:"Mechanisms and Adaptations",isOpenForSubmission:!1,hash:"588466f487e307619849d72389178a74",slug:"abiotic-stress-in-plants-mechanisms-and-adaptations",bookSignature:"Arun Shanker and B. 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Recommendations concerning using feed additives, their categories and the description of requirements related to such additives can be found in the regulation (EU) No 1831/2003 [1]. Detailed regulations oblige the entities launching feed additives on the market to specify the methods used to analyse active substances of additives for the needs of official feed control. Official feed control is implemented in order to monitor adequate and safe use of feed additives in animal nutrition. Moreover, controlling feed production in this respect results in improving the quality and safety of animal products for consumers.
HPLC methods have been widely used in the analyses of feed additives, such as vitamins, feed colorants, antioxidants, amino acids and coccidiostats in preparations, premixes and feed mixtures. It is relatively simple to analyse preparations as they are usually composed of a particular active substance and a carrier. A premix is more complex feed consisting of a combination of a dozen different feed additives on a mineral (calcium carbonate) or organic (wheat bran) carrier. Complete feed mixtures used in animal nutrition, produced by combining premixes with feed materials are often greased and subjected to further hydro- or barothermal processing, e.g. pelleting, extrusion or expanding. In order to counteract decomposition of the active substance, feed additives are secured by protective coating, e.g. vitamin A, canthaxanthin, which enhances their durability in the feed matrix. A specific protection of feed additives by protective coating, thermal processing, greasing the feed, varied composition of feed materials in mixtures may hinder the transfer of the active substance into a solution during extraction and purification of the extract.
The key issue becomes selection of a chromatographic column (in a normal phase or reversed phase), mobile phase, detector, as well as optimization the conditions of chromatographic separation. In examining feed additives with the use of HPLC methods the most frequently used types are spectrophotometric detection (UV-VIS), detection with the help of diode array and fluorescent detection. The choice of optimal parameters for chromatographic separation is done during validation of the method. The analyses in this respect should be accompanied by an assessment of the method’s robustness [2]. A practical way to verify the precision of a method in a laboratory (repeatability) and in interlaboratory studies (reproducibility) is taking advantage of Horwitz equation [3].
Using HPLC methods for examining feed additives was the subject matter of numerous studies on the basis of which official methods of analysing certain feed additives were developed. The studies presented the basic validation parameters for the methods of examining the content of fat-soluble vitamins [4-7], water-soluble vitamins [8,9], coccidiostats [4,10-12], and other feed additives, amino acids, methionine hydroxy analog and antioxidants [13,4,8]. However, in case of carotenoids such as canthaxanthin or apocarotenoic acid ester official methods of examining these additives are still based on spectrophotometric measurement rather than on HPLC methods [13,8].
New requirements have been introduced regarding the validation parameters for the methods of analysing feed additives, e.g. those listed in the regulation No 882/2004 [14], taking into consideration, among others, the uncertainty of measurement. It is necessary to determine the uncertainty of measurement with a particular method in order to interpret adequately the result of examining feed additives in feedingstuffs and to assess acceptable tolerance in compliance with the requirements of the regulation No 939/2010 [15]. The new requirements in this area should be taken into account while validating the methods of testing feed additives in order to solve the problem of interpreting the results.
The aim of the present work was to offer a review of HPLC methods used for analysing active substances in certain feed additives, with regard to current requirements defined in the regulations. In some justified cases the results of the authors’ own studies were presented, as well as the procedures for determining vitamins B1 and B2, canthaxanthin and methionine hydroxy analog (MHA). Some validation parameters were presented, such as the limit of the method’s quantification (LOQ), linearity of the calibration curve, repeatability, within-laboratory reproducibility (intermediate precision), recovery and the uncertainty of measurement. Also, the results of verifying the developed methods and laboratories participating in proficiency testing (PT) were demonstrated. The ways of quality assurance of the tests in reference to HPLC methods were discussed. The work presents the method of assessing combined standard uncertainty of measurement with the use of experimental approaches based on within-laboratory reproducibility and calculations for the bias of the method on the basis of CRM studies or PT results.
Feed additives are commonly used in animal nutrition, e.g. in order to supplement the animals’ requirement for nutrients (amino acids), useful micro components (vitamins), to prevent invasive diseases (e.g. coccidiosis – coccidiostats), to reduce oxidation processes of feed’s components (antioxidants), to enhance the dietary value and quality in food products of animal origin (amino acids, carotenoids– egg yolk coloration).
In case of some vitamins and antioxidants their maximum content in feed mixtures was determined, e.g. for vitamins A and D3, antioxidants (ethoxyquin, BHA, BHT), carotenoids (canthaxanthin, apocarotenoic acid ester and others). Maximum contents are subject to official control in reference to their conformity with the requirements related to the safety of feedingstuffs. Additionally, a feed manufacturer is obliged to declare the content of feed additives on the label of a premix or feed mixture. Thus, it is necessary to have access to analytical methods for testing the content of feed additives in a wide range of concentrations in the preparations containing additives, premixes and feed mixtures.
Table 1 presents examples of well-known HPLC methods for examining fat-soluble vitamins in feedingstuffs, including the official methods accepted by the European Commission. A commonly used method of preparing a sample for analysing the content of vitamin A is alkaline hydrolysis during which gelatin/sugar cross-linked beadlets which protect vitamin A in the form of retinol acetate are solved and then purified by liquid-liquid extraction. An interesting option for purifying vitamin A extracts from feed mixture with the use of the SPE technique was presented by Fedder & Ploger [7]. The step of alkaline hydrolysis is also used while determining vitamins E and D3. Chromatographic separation of vitamins, except for vitamin D3 where preliminary separation and fraction collection are necessary [5], does not present any serious problems. However, a problem may be posed by the quality and durability of a standard, as well as poor precision resulting from a too low of analytical weight [16]. It is necessary to verify vitamin A standards with the use of a spectrophotometric method [4].
The official methods of determining the content of water-soluble vitamins, such as B1, B2 and B6 are based on spectrophotometric or fluorometric methods [8,13]. The results of analyses using these methods may be biased with errors due to some interferences from other substances in the variable feed matrix. Recently HPLC methods to determine vitamins B1, B2, B6, nicotinic acid and nicotinamide in mineral preparations and mixtures [8], as well as vitamin B1 in feed mixtures and premixes [9] were published (Table 2). Due to the high limit of quantification for vitamin B1 amounting to 5 mg/kg according to Italian Official Method [9], it cannot be used for analysing vitamin B1 in typical feed mixtures to which it is normally added at the amount of 2-4 mg/kg. Moreover, the method quoted above makes it possible to examine vitamin B1 added to feedingstuffs but not the total content of this vitamin, regarding its presence in feed materials. It is thus necessary to have access to chromatographic methods enabling the examination of water-soluble vitamins present in feed materials and added in the form of feed additives. The procedures of HPLC methods of vitamins B1 and B2 developed during the authors’ own studies are presented later in the chapter [17,18].
Analyte, matrix reference | Extraction, extract clean up and chromatography parameters | Performance parameters |
Vitamin A in feedingstuffs and premixes, Commission Regulation, 2009 [4] | Sample hydrolyze with ethanolic KOH, extraction into light petroleum, evaporation and dissolution in methanol, reversed phase HPLC, C18 column (250 x 4 mm) 5 μm or 10 μm packing , mobile phase: methanol and water 98+2 (v/v), fluorescence (or UV) detector: excitation 325 nm; emission 475 nm or UV detector (325 nm) | LOQ=2000 IU/kg; SDr (%): 3.0-8.1; SDR (%): 6.2-20.0; |
Vitamin E in feedingstuffs and premixes, Commission Regulation, 2009 [4] | Sample hydrolyze with ethanolic KOH solution, extraction into light petroleum, evaporation and dissolution in methanol, reversed phase HPLC, C18 column (250 x 4 mm) 5 μm or 10 μm packing , mobile phase: methanol and water 98+2 (v/v), fluorescence detector: excitation 295 nm; emission 330 nm or UV detector (292 nm) | LOD=2 mg/kg; LOQ=10 mg/kg; SDr (%): 2.2-4.1; SDR (%): 4.8-12.7 |
Vitamin D3 in feedingstuffs and premixes [5] | Feed saponification and extraction with diethyl ether; evaporation and solvation in methanol; reverse phase preparative chromatography; eluat collection with vitamin D3 , evaporation and next solvation in n-hexan or isooctane; normal phase chromatography, column 250 mm x 4 mm, Si-60, 5 μm packing; UV detection at 264 nm; mobile phase: preparative column: methanol -water (92+8)), analytical column – n-hexan – dioxan - isopropanol (94.5+5+0.5) | LOQ=1000 IU/kg; RSDr to 5000 IU/kg: 1000 IU/kg; 5000-20000 IU/kg:20% 20000-100000 IU/kg: 15%; "/>100000 IU/kg: 10% |
Vitamin K3 in feedingstuffs, premixes and feed additives [6] | Sample extraction with chloroform, transfer vitamin K substances to free menadion; clean-up with Celite and sodium sulphate anhydrous; normal phase chromatography, Si-60 column 250 mm x 4 mm, 10 μm packing; UV detection at 251 nm | LOQ=0.5 mg/kg RSDr at 1 mg/kg: 10%; SDr at 8 mg/kg: 4%; SDr at the level "/>2500 mg/kg: 3%; |
Vitamin A and E, feedingstuffs [7] | Sample hydrolyze with ethanolic KOH solution; clean-up on SPE column; elution in ethyl acetate, evaporation and dissolution in methanol; reverse phase chromatography, ODS2 column 250 mm x 4.6 mm, 5 μm packing; UV detection at 325 nm (vitamin A) and 292 nm (vitamin E). | Range: Vitamin A=1250-20000 U/kg; Vitamin E=3-300 mg/kg RSDip= 21% (vit. A), 11% vit. E; Rec: vit. A - 80%, vit. E - 110% |
HPLC methods for the analysis of fat-soluble vitamins in feeds
Analyte, matrix reference | Extraction, extract clean up and chromatography parameters | Performance Parameters |
Vitamin B1, B2, B6, NA, NSA in premixes and mineral feeds [8] | Extraction with methanol -Titriplex solution, clean-up on membrane filter 0.45 μm, reverse phase HPLC coupled to UV or diode array detector, column Nucleosil 250 x 3.0 mm, 5 μm packing; mobile phase: mixture of water solution of acetonitrile and acetic acid | Range, mg/kg: B1= 320-7940; B2= 868-15990; B6=627-11530; NA=4520-77850; NSA= 3665-61230; RSDr(%) = 2.1-5.1; RSDR(%) = 4.2-30.2 |
Vitamin B1 in feedingstuffs and premixes [9] | Extraction with methanol ; clean-up on SPE, reverse phase HPLC, coupled to a fluorescence detector, excitation at 360 nm, emission at 430 nm | LOQ=5 mg/kg; Range, mg/kg: 7 – 484; RSDr(%)=4.2-4.7 RSDR(%)=5-13 Rec.(%)=88-97 |
HPLC methods for the analysis of water-soluble vitamins in feeds
Numerous methods have been developed to examine coccidiostats in feedingstuffs with the use of high performance liquid chromatography. Examples of such methods are presented in Table 3. Satisfactory precision of such methods has been obtained, in conformity with that calculated from the Horwitz equation [3] and with the requirements of the Commission’s Decision [19], which enables analyzing coccidiostats at the levels declared by manufacturers. Due to the hazard of cross-contamination with the remains of coccidiostats found in non-target mixed feeds on the production line and the risk of carry-over the remains of contamination onto the products of animal origin, it is necessary to continue lowering the limit of methods’ quantification in order to control safe use of coccidiostats.
Table 4 presents HPLC methods for testing other feed additives, such as antioxidants, amino acids, methionine hydroxy analog. The official AOAC method for determining ethoxyquin was verified in testing pet food and meat meal [13]. Due to the determination of maximum content of antioxidants in feed mixtures for animals used for food production there is a necessity to check this method in testing typical feed mixtures and premixes. Amino acids are present in typical feed materials as components of proteins. In order to determine amino acids in feed materials it is necessary to subject proteins to hydrolysis and next to separate amino acids using ion-exchange chromatography and apply derivatization. With intensive animal production it is necessary to supplement the deficiency of amino acids, such as lysine, methionine, threonine and tryptophan. New feed additives have been registered recently, such as arginine, valine and cysteine. The official AOAC methods make it possible to determine mainly the composition and content of amino acids in feedingstuffs after hydrolysis [13], yet validation parameters of the determination methods have not been defined for all synthetic amino acids. Moreover, the precision parameters of the method used to determine amino acids with sodium metabisulphite or the hydrobromic acid method were in many cases unsatisfactory, which was confirmed by the values of the Horwitz ratio higher than 2, e.g. in a feed mixture for broiler chickens the HorRat (H) values amounted to 1.7-3.6, with mean 2.5, while satisfactory H values are within the range of 0.5 > H > 2. This requires further studies with the use of high performance liquid chromatography in order to determine the total content of amino acids after hydrolysis and added amino acids.
Analyte, matrix reference | Extraction, extract clean up and chromatography parameters | Performance Parameters |
Halofuginone, medicated feeds [10,4] | Ethyl acetate extraction, purification by ion-exchange chromatography, reversed phase HPLC with UV detection at 243 nm, C18 column (300 x 10 mm) 10 μm packing, mobile phase: mixture of acetonitrile and ammonium acetate buffer solution | LOQ = 1 mg/kg ; RSDr (%): 2.0-4.7; Rec.(%) 75.3-98.0; at the level of 3 mg/kg |
Lasalocid, monensin, salinomycin and narasin, poultry feed [12] | Methanol extraction without clean-up, derivatization with 2,4-dinitrophenylhydrazide (DNP) in acid medium at 55 °C, ODS column (150 x 4.6 mm, 5 μm); eluent: methanol – 1.5% aqueous acetic acid (90:10, v/v), UV detection at 305/392 nm | LOQ = 40 mg/kg conc. range 50-150 mg/kg; RSDr (%): 4-10; Rec. 85-100% |
Lasalocid, poultry feeds, premixes [11,4] | Extraction into acidified (HCl) methanol, agitation in ultrasonic bath at 40 ºC, filtration through a 0.45 μm filter, reversed phase HPLC, C18 column (125 x 4 mm) 5 μm packing , mobile phase: mixture of phosphorus buffer solution and methanol 5+95 (v/v), fluorescence detector: excitation 310 nm; emission 419 nm | LOD=5 mg/kg; LOQ=10 mg/kg; RSDr (%): 2.1-5.4; RSDR (%): 5.0-10.7; Rec : feed ≥ 80%; premixes ≥ 90% |
Robenidine, feedingstuffs, premixes [4] | Extraction into acidified (HCl) methanol, clean-up on an aluminum oxide column; reversed phase HPLC, UV detection at 317 nm; C18 column (300 x 4 mm) 10 μm packing; mobile phase: mixture of acetonitrile and sodium and potassium phosphate solution | LOQ=5 mg/kg SDr (%): 3.3-5.4; SDR (%): 9.7-10.1; Rec. for blanc sample ≥ 85% |
Diclazuril, feedingstuffs, premixes [4] | Extraction with acidified methanol with internal standard; purification on C18 solid phase extraction cartridge (SPE), evaporation and dissolution in DMF; reversed phase gradient HPLC, Hypersil ODS column, 100 mm x 4.6 mm, 3 μm packing; mobile phase: (1) aqueous solution of ammonium acetate and tetrabutyl-ammonium hydrogen sulphate, (2) acetonitrile, (3) methanol | LOD=0.1 mg/kg; LOQ=0.5mg/kg; SDr (%): 1.9-17.3; SDR (%): 7.4-18.6; Rec. for blanc sample ≥ 80% |
HPLC methods for the analysis of coccidiostats in feeds
Analyte, matrix reference | Extraction, extract clean up and chromatography parameters | Performance Parameters |
Ethoxyquin in pet-food and meat meal [13] | Extraction with acetonitrile without clean-up, reversed phase HPLC, C18 column (250 x 4.6 mm) 5 μm packing , mobile phase: acetonitrile and 0.01 M ammonium acetate (70 + 30, v/v); fluorescence detector: excitation 360 nm; emission 432 | Method range: 0.5-300 mg/kg; SDr (%): 4.5-32; SDR (%): 4.5-55; Rec. 60-83% |
Phenolic anti-oxidants* in fats [13] | Extraction with acetonitrile, extract is concentrated and diluted with 2-propanol; reversed phase gradient HPLC, C18 column with guard column; mobile phase: (1) 5% acetic acid in water, (2) acetonitrile-methanol (1 +1, v/v) | Method range: 10-200 mg/kg; SDr (%): 2.1-11.5; SDR (%): 2.7-21.5; Rec. 83-103% |
Tryptophan in feedingstuffs and premixes[4] | For total tryptophan alkaline hydrolise with saturated barium hydroxide solution; for free tryptophan extraction under mild acid conditions; reversed phase HPLC with fluorescence detector, excitation 280 nm, emission 356 nm; C18 column (125 x 4 mm) 3 μm packing; mobile phase: acetic acid and 1,1,1-trichloro-2-methyl-2-propanol solution, pH 5.00 | Feedingstuffs: SDr (%): 1.6-1.9; SDR (%): 2.2-6.3; Feed materials: SDr (%): 0.8-1.3; SDR (%): 4.1-5.1; |
Amino acids in feeds** [13] | Performic acid oxidation of the sample to oxidize cystine and methionine; amino acids liberation from protein by hydrolysis with 6 M HCl; dilution with sodium citrate buffer; amino acid separation on ion-exchange chromatograph with ninhydrin post-column derivatisation | Broiler feed: SDr (%): 1.1-4.7; SDR (%): 6.0-19.8; HorRat: 1.7-3.6 ~ 2.5 |
MHA in feedingstaffs and premixes [8] | Extraction with water solution of acetonitrile; reversed phase HPLC with UV detection at 210 nm; RoSil-NH2 column (250 mm x 4.6 mm, 5 μm packing) with guard column; mobile phase: acetonitrile with phosphoric acid solution ( 23+77) | LOD=0.2 g/kg LOQ=0.5 g/kg |
HPLC methods for the analysis of other feed additives in feeds
The difficulty in determining certain feed additives is related with their low stability. In order to obtain a more durable form, resistant to the manufacturing conditions of feed mixtures, the additives are secured by protective coating. This concerns primarily vitamins A and D3, as well as feed colorants, such as canthaxanthin.
Vitamin A is produced in the form of gelatin-and-sugar beadlets or fat beadlets. Each beadlet contains ca. 0.5-0.6 µg of vitamin A, as calculated for retinal (ca. 2 IU). The distribution of beadlets in the feed is not equal and the feed enriched in vitamin A tends to segregate vitamin beadlets during the process of manufacturing and transporting the feedingstuff, especially in case of loose products. On the other hand, pelleting feed mixtures or subjecting them to other barothermal processes, such as extrusion or expanding reduces vitamin segregation, yet it lowers their durability at the same time. Ultimately, the unequal distribution of vitamins in feed may affect the precision and accuracy of results of analyses. Grinding the samples may improve the distribution of vitamin A, yet it will also increase the risk of its oxidation. Vitamin A is chemically unstable and its content and biological activity are reduced along with the presence of oxygen from the air, light, humidity, inorganic acids, choline hydrochloride, microelements and peroxides created in the processes of fat oxidation. It is recommended that samples should be ground immediately prior to an analysis into 1 mm particles. Further grinding of the sample before determining the content of vitamins may lead to their decomposition. A useful guideline regarding the preparation of samples for analyses, including the analyses of unstable feed additives such as vitamins, is provided by the currently issued ISO/FDIS International Standard 6498 [20].
Vitamins are protected against oxidation by antioxidants (e.g. ethoxyquin) which are added to the materials creating beadlets. Some forms which are physically and chemically stable are created in this way, e.g. the oleic form of vitamin D3 and more stable compounds (menadione bisulfite, thiamine mononitrate and riboflavin phosphate). Feed additives used in feed manufacturing are introduced on mineral carriers or on wheat bran. The most frequently used mineral carrier is fodder chalk. In case of extracting additives from premixes with the use of diluted acids the influence of the carrier on the conditions of extraction should be considered. In such a situation it is recommended that the robustness of a method to slight changes in the analytical procedure or a change of matrix should be verified [2]. In analyzing feedingstuffs the interfering agent is fat which often occurs in significant amounts on feed mixtures (up to 10%). In high-fat samples containing more than 0.25g of fat in an analytical weight while determining fat-soluble vitamins, additional soaps are formed in the saponification process, which hinder the separation of the examined analyte.
It was possible to resolve the problem of interfering substances after using HPLC methods. However, the diversity of matrices and inhomogeneity of feedingstuffs pose numerous analytical problems while determining feed additives, such as vitamins or carotenoid colorants. The biggest difficulty is related to proper clean-up of the extract and selection of adequate conditions for chromatographic separation.
In case of vitamins, while examining the relevance of the producer’s declaration and interpreting the result of the examination, one should be aware of the effect brought about by numerous factors on the content of vitamins in feedingstuffs. There were detailed studies in this respect conducted by Coelho [21]. Table 5, based on Coelho’s article, presents the method of estimating the summary influence of different factors, such as the type of a premix and the time passed since the moment of its production (column 2), the type of conditions of hydro- and barothermal processing (column 3), feed storage time (column 4) on vitamin retention. The product of the factors in columns 2, 3 and 4 (expressed as a fraction) will let us estimate the retention of a particular vitamin in a particular feed (column 5).
Vitamin | Vitamin, premix (Coelho [21], Table 8) | Pelleting temp., (Coelho[21], Table 11) | Feed storage time (Coelho [21], Table 10) | Total vita-min reten. % |
1 | 2 | 3 | 4 | 5 |
2 months | 96 °C | 2 weeks | 2 x 3 x 4 | |
A beadlet | 90 | 88 | 98 | 78 |
D3 beadlet | 91 | 91 | 99 | 82 |
E acetate 50% | 92 | 91 | 99 | 83 |
Thiamine | 77 | 82 | 99 | 63 |
Riboflavin | 91 | 84 | 99 | 76 |
B12 | 96 | 95 | 100 | 90 |
Ca pantothenate | 87 | 84 | 99 | 72 |
Biotin | 89 | 84 | 99 | 74 |
Niacin | 90 | 86 | 99 | 77 |
Vitamin stability in premixes and feeds (%), (Coelho, 2002)
Similarly, the authors observed in their own studies that the conditions and premix storage time in the laboratory affected the content of vitamins A and E. Fractioned samples of premix, each weighing 100 g, were stored for 8 months at room temperature (22 °C), in a fridge (5 °C) and in a freezer (-18 °C) (Table 6). In the samples stored at room temperature the content of vitamins after 8 months of storage was reduced to as little as 3% of the initial value. The analyses of vitamin content should be performed immediately after receiving the samples by the laboratory, otherwise the samples should be stored in a fridge until the analyses can be done.
Item | Vitamin retention, %* | ||||||||
Month | |||||||||
0 | 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | |
Vitamin A, initial value 2794000 IU/kg: - room 22 °C - refrigerator 5 °C - freezer -18 °C | 100 100 100 | 94 102 100 | 82 99 100 | 64 94 100 | 32 94 99 | 13 93 100 | 6 93 98 | 4 92 99 | 3 85 96 |
Vitamin E, initial value 15.56 g/kg: - room 22 °C - refrigerator 5 °C - freezer -18 °C | 100 100 100 | 98 99 99 | 96 98 97 | 94 100 100 | 94 98 99 | 97 100 100 | 95 100 100 | 94 98 100 | 90 90 94 |
The results of laboratory retention of vitamins A and E, %
A sample for a quantitative analysis should be prepared in such a way that the isolation of a selected analyte and removal of interfering substances is possible. The condition necessary for adequate quantitative determination in liquid chromatography is eliminating any possibility of coelution and minimization the drift of a basic line. Extractions of the analyzed substances were done classically by shaking the sample with a solvent. In case of vitamins B1 and B2 and carotenoids, such as canthaxanthin and apocarotenic acid ester, extraction was performed in an ultrasonic bath. In order to purify the extract, aluminum oxide (e.g. canthaxanthin), celite and anhydrous sodium sulphate (e.g. vitamin K3) were used and PTFE and Nylon (PA) (0.45 µm or 0.20 µm) syringe filters were applied before injecting on the chromatographic column. Syringe (hydrophobic) PTFE filters are used in case of solutions with high acid and base content, whereas nylon (hydrophilic) filters are used with aqueous and organic solutions. Filtration of extracts is necessary as it prolongs durability of a column due to eliminating permanent contamination which blocks the column’s intake and increases back pressure. If needed, the analyte may be concentrated by evaporating the solvent. When there is a risk that the studied analyte becomes oxidized, evaporation is done in neutral gas, e.g. nitrogen or argon.
The removal of gelatin-and-sugar beadlets protecting vitamins A and D3 is done at the stage of saponification and transforming the vitamins into alcoholic forms. Douša & Břicháč [16] demonstrated that saponification in standard conditions did not affect the results of analyses. In case of canthaxanthin enzymatic hydrolysis through adding trypsin and pepsin is used. While determining the total content of thiamine and/or riboflavin in feedingstuffs (endogenic and added) at the stage of preliminary preparation an ultrasonic bath and also enzymatic hydrolysis (taka-diastase) are applied.
The method of high performance liquid chromatography (HPLC) with flouorometric detection or a diode array is characterized by sufficient selectivity and sensitivity required to determine feed additives. Each of the developed and verified procedures includes precisely defined stages of hydrolysis and extraction which make it possible to determine the total or added analyte in a sample. Chromatographic separation of the analyzed mixture is affected by the properties of chromatographic arrangement. The developed analytical methods took advantage of chromatography in a regular and reversed arrangement of phases. In the reversed phase methanol and water or acetonitrile and water were used. In the standard arrangement the mobile phase was hexane or chloroform. In the majority of the developed methods presented in this chapter isocratic elution was applied, except for methionine hydroxy analog where gradient elution was used. Adequate separation was achieved through the use of ODS column with 18 atoms of carbon in the chain (C18) and columns with 8 atoms of carbon (C8) in the alkyl chain in chromatography with reversed phase (RP). In chromatography with the normal phase arrangement columns filled with silica gel were used. The identification and content of analyte was examined with the method of absolute calibration (with external model), analyzing separately the sample and the model and identifying the peaks with the help of retention values, comparing retention time of the identified substance with the retention time of a standard, chromatographed in identical conditions. Examples of chromatograms for the standard extract and the sample of the examined analyte (riboflavin) with the use of fluorescent detector are presented below (Fig.1, Fig.2).
Characteristic chromatograms of riboflavin: chromatogram of standard solution
Characteristic chromatograms of riboflavin: chromatogram of premix extract
Selected methods of testing feed additives presented below were validated with the help of a high pressure liquid chromatograph (Dionex P-680) with fluorescence detector (Dionex RF 2000) or with a diode array.
The following procedure used for determination of thiamine in premixes and compound feeds was elaborated on the basis of the article published by Rubaj
Vitamin B1 is extracted with hydrochloric acid of 0.1 mol/l and next oxidized to thiochrome and marked with the use of high performance liquid chromatography (HPLC) with a fluorescence detection.
Thiamine was extracted from the examined feed sample with 0.1 M hydrochloric acid at 100°C for 30 minutes. In case of compound feedingstuffs 10% taka-diastase solution was added to the samples, and then samples were incubated at 37°C for 17 hours. Afterwards thiamine was oxidized to thiochrom by 1% alkaline K3Fe(CN)6.
Column 25 cm x 4.6 mm
Stationary phaseLichroCart 250-4, Lichrospher100 NH2 (5µm)
Mobile phaseChloroform and methanol, 90+10 (v/v)
Column temperature25 ºC
Flow rate2.0 ml/min
Injection20 µl
DetectorFluorescence, Ex λ= 365, Em λ=435
CalculationExternal standard, peak area, linear regression
This method was applied for the quantification of total content of thiamine in compound feedingstuffs as well as added thiamine in the form of hydrochloric or nitrate salt.
The following procedure for determination of riboflavin in premixes and compound feeds was elaborated on the basis of the article published by Rubaj et al. [17].
Riboflavin was extracted from a feed sample in autoclave with 0.1M sulphuric acid. The ester bonds with phosphoric acid were hydrolyzed by the Taka-diastase enzyme. Riboflavin content was determined by high performance liquid chromatography (HPLC) with reversed-phase and usage of fluorescence detection.
Riboflavin was extracted from the examined feed sample with 0.1M sulphuric acid, and that solution was boiled for 15 min. at temperature from 110°C to 120°C. After cooling to the room temperature, the whole volume of hydrolysed sample was transferred to a 100 ml measuring flask. Next taka-diastase suspension was added to the flask, which was then placed into a water bath at 45°C for 20 min. The enzymatic reaction was stopped by adding sulphuric acid. The sample solution was next chilled to room temperature, and the volume was corrected to 100 ml by adding 0.1 mol/l sulphuric acid. Afterwards, samples were mixed and filtrated. Extract clean-up was done by adding methanol to the sample and filtration through syringe filter before injection on the column.
Column 25 cm x 4.6 mm
Stationary phaseC18
Mobile phaseMethanol and citric acid 0.2 mg/l (30:70 v/v). That solution was mixed with methanol with ratio 1:1
Column temperature25 ºC
Flow rate0.8 ml/min
Injection20 µl
DetectorFluorescence, Ex λ= 453, Em λ=521
CalculationExternal standard, peak area, linear regression
Vitamin B2 is sensitive to light, hence all the activities were conducted without any access of day light (by using amber glass flask or flask covered by aluminum foil).
The following procedure for determination of canthaxanthin in premixes and compound feeds was elaborated on the basis of the article published by Rubaj et al. [22].
The principle of this method is based on the hydrolysis of a powdered formulation of canthaxanthine with trypsin and pepsin in water solution of ammonia and its purification on the aluminium oxide filled column. The canthaxantine content is determined by high performance liquid chromatography (HPLC) in normal phase with usage of DAD detector.
The sample is hydrolyzed with an aqueous solution of ammonia at the presence of trypsin and pepsin following extraction with ethyl alcohol and diethyl ether. Purification occurs on the aluminum oxide filled column. The extract prepared in this way should be evaporated, dissolved in the mobile phase, filtered through syringe filters and dosed on the column.
Column 4.6 x 250 mm
Stationary phaseLiChrospher Si 60
Mobile phasen-hexane: acetone 86:14 (v/v)
Column temperature25 ºC
Flow rate1.3 ml/min
Injection20 µl
DetectorDAD λ=446 nm
CalculationExternal standard, peak area, linear regression
The procedure of analyzing methionine hydroxy analog was developed on the basis of the work by Matyka
Methionine hydroxy analog is extracted from the sample by 10% acetonitrile, and next hydrolyzed with potassium hydroxide and determined by high performance liquid chromatography (HPLC) with reversed phase and UV detection.
Extract methionine hydroxy analog from the feed, with the use of 10% acetonitrile. After centrifuging, perform hydrolysis with potassium hydroxide and next with a solution of orthophosphoric acid. Filter the supernatant through syringe filters and inject on the column.
Column 25 cm x 4.6 mm
Stationary phaseC18, LiChrospher
Mobile phaseEluent 1 : acetonitrile - phosphoric acid 10+90 (v/v)
Eluent 2 : acetonitrile - phosphoric acid 23+77 (v/v)
Column temperature25 °C
Flow rate0.8 ml/min
Injection20 µl
DetectorUV, λ=214 nm
CalculationExternal standard, peak area, linear regression
If the degree of MHA polymerization in the feed mixture is high and depolymerization in the conditions presented in the analytical procedure is incomplete it is necessary to increase the amount of hydroxide taken for hydrolysis and the amount of phosphoric acid for neutralization, keeping constant proportions, and next to take into account the change in the amount of the solution after hydrolysis, while calculating MHA content.
The quality of separation on the chromatographic column depends on pH of the mobile phase. When acidity increases retention time for MHA is reduced. The excess of phosphoric acid in the injected solution after depolymerization reduces the time of MHA retention.
A significant element in verifying a chemical method, including chromatography, is its validation. Validation is a confirmation through examining and presenting some objective evidence that some particular requirements regarding the intended application have been fulfilled. The basic validation parameters include: calibration linearity, the limit of detection and quantitative determination, precision (repeatability, indirect precision, reproducibility), recovery and uncertainty.
Calibration linearity is defined as a relationship between the readings of the measuring device and the concentration of a particular component, in conformity with the regression equation: bx + a = y. The measure of linearity is Pearson’s linear correlation coefficient (r) for parameters with regular distribution. The scale presented below is adopted to estimate the correlation coefficient: 0.0-0.2: very weak relationship; 0.2-0.4: weak relationship; 0.4-0.6: moderate relationship; 0.6-0.8: strong relationship; 0.8-1.0: very strong relationship.
In case of feed additives discussed in the present chapter, determined with HPLC methods, external calibration was used.
In case of chromatographic methods the value of limit of detection (LOD) may be determined on the basis of the obtained chromatogram of blanc sample, as the threefold value of a noise signal. To do this, it is necessary to determine the level of noise, by measuring on the chromatogram the range of signal change near retention time of examined analyte. With chromatographic methods, the bottom limit of the method’s application may be also regarded as the content of the analyzed component, which is equal to the lowest concentration of the standard used for calibration.
Analyte | Matrix | LOQ | CVr % | CVip % | Rec. % | Linear range |
Vitamin A | Feedingstuff | 1000 IU/kg | 1.6 | 4.0 | 96.0 | 7.0-70 IU/ml; r=0.999 |
Premixture | 1.4 | 2.0 | 95.2 | |||
Vitamin E | Feedingstuff | 6.0 mg/kg | 2.0 | 2.0 | 96.7 | 0.05-0.3 mg/ml; r=0.999 |
Premixture | 1.0 | 2.0 | 96.4 | |||
Vitamin K3 | Feedingstuff | 1.0 mg/kg | 6.4 | - | 100.9 | 0.046-4.62 µg/ml; r=0.999 |
Premixture | 5.7 | - | 99.4 | |||
Preparation | 1.9 | - | 101.2 | |||
Vitamin D3 | Premixture | 200 IU/g | 1.4 | 1.7 | 99.3 | 1.06-10.68 µg/ml; r=0.999 |
Preparation | 1.3 | 1.3 | 98.4 | |||
Vitamin B1 | Feedingstuff | 1.0 mg/kg | 5.6 | 98.9 | 0.2-1.0 µg/ml; r=0.999 | |
Premixture | 3.7 | 102.3 | ||||
Vitamin B2 | Feedingstuff | 1.0 mg/kg | 3.4 | 5.1 | 98.0 | 0.17-0.67 µg/ml; r=0.999 |
Premixture | 2.3 | 6.2 | 98.3 |
Validation parameters obtained for selected feed additives – vitamins in feeds
Analyte | Matrix | LOQ | CVr % | CVip % | Rec. % | Linear range |
Cantha-xanthin | Feedingstuff | 1.0 mg/kg | 4.7 | 7.9 | 97.3 | 0.7-8.5 µg/ml; r=0.999 |
Premixture | 3.3 | 6.0 | 98.2 | |||
Tryptophan | Feedingstuff | 10.0 mg/kg | 4.1 | 4.0 | 94.9 | 12.5-100 nmol/l; r=0.999 |
Preparation | 1.0 | 1.4 | 99.7 | |||
Ethoxyquin | Feedingstuff | 0.5 mg/kg | 2.0 | 6.0 | 99.0 | 0.01-0.07 µg/ml; r=0.999 |
MHA | Feedingstuff | 0.05% | 2.8 | - | 96.7 | 0.05-0.45 mg/ml; r=0.997 |
Validation parameters obtained for other feeds
During the validation process in a laboratory the precision of a method is determined through examining such parameters as repeatability and within-laboratory reproducibility (intermediate precision). Within-laboratory reproducibility may be calculated on the basis of control charts or from the range between parallel results of an analysis (replications) of a feed additive, in compliance with the Nordtest [23] handbook. For two or more replications for the analyses of an analyte in each sample it is necessary to calculate the mean value, the difference between measurements (range), relative difference in % and next mean relative difference (%) for all samples of a particular type of feed. The mean range divided by the coefficient (for two replications d = 1.128) makes the standard deviation of within -laboratory reproducibility. In order to verify the method’s precision the Horwitz ratio named HorRat (H), may be used which is the ratio of the relative standard deviation of reproducibility SDRt calculated from the Horwitz formula SDRt =2 C-0.15, where C stands for concentration expressed as a dimentionless mass fraction (e.g. 1 mg/kg = 10-6). In order to adjust it to the conditions of repeatability, target standard deviation SDRt is multiplied by 0.50 (RSDr = 0.5 RSDR), [3]. Satisfactory values of the HorRat making the measurement of precision are included in the range of 0.5 < H < 2 [3]. In case of participating in interlaboratory tests and obtaining satisfactory results, it is possible to include the precision parameters obtained in these analyses. The accuracy of a method may be determined by calculating the recovery degree or examining certified reference material, CRM.
Each laboratory should possess a program of quality assurance of its analyses within good laboratory practice. In case of chromatographic methods steering the quality may be implemented through performing one or more of the activities listed below:
regular examinations of control samples;
regular check-ups of the standard for each examination or series of analyses of labile feed components,
checking b curve slope coefficient from the equation bx + a=y,
analyzing overlapping samples (e.g. a solution of the sample for measurement, prepared and analyzed on the previous day),
analyzing the blanc sample and fortified sample,
examining certified reference materials, reference materials,
participation in native and international proficiency testing.
Control material may be provided by certified reference material, CRM (matrix + analyzed substance), material from proficiency testing with a value assigned, enriched material prepared in the laboratory (fortified sample) and control material with recognized content of the tested and stable in time component, previously determined in the laboratory.
In compliance with the recommendations of the EN ISO/IEC 17025:2005 [24] standard and requirements defined in some regulations, in order to assess and interpret the result of a test, it is necessary to use the uncertainty of measurement. We hardly ever know the real content of the analyte and the result of the test is biased with an error. Hence, the idea of “uncertainty of measurement” has been introduced which is defined as “a parameter associated with the result of a measurement, that characterises the dispersion of the values that could reasonably be attributed to the measurand” [26]. The EN ISO/IEC 17025:2005 standard recommends at point 5.4.6.2 that testing laboratories should possess and make use of procedures for assessing the uncertainty of measurement. To assess the uncertainty of methods used to analyze feeds the most frequently used approach is the model, consistent with the GUM [26] guidebook, which consists of finding the components of uncertainty and uses the law of error propagation. Using this particular approach to assess uncertainty, it is possible to obtain an underestimated value in case when we do not consider all the components. Other reasons for underestimating uncertainty during validation include a situation when while assessing uncertainty repeatability instead of within-laboratory reproducibility is taken into account or we often forget to consider the bias. Moreover, uncertainty assessment is done during validation when well-ground typical samples are analyzed in a short time, new standards are prepared, the apparatus is controlled (standard conditions). During routine activities we analyze various matrices and obtaining homogeneity is not easy in case of some samples. The conditions mentioned above may affect underestimation of uncertainty. That is why it is important to have a possibility to verify uncertainty with the help of other approaches.
New opportunities concerning verification and assessment of uncertainty can be found in the Eurolab [27] technical report, the Nordtest [23] handbook and in the paper [28] which recommend the use of experimental approaches in assessing uncertainty of laboratory methods, in particular:
within-laboratory experimental approach based on within -laboratory reproducibility and the assessment of the method’s bias, following CRM,
within -laboratory experimental approach based on within -laboratory reproducibility and the assessment of the method’s bias, following participation in PT/ILC.
Using the within-laboratory experimental approach in assessing uncertainty of measurement (u) within -laboratory reproducibility (s) is considered as a measure of precision, as well as the bias (b), in accordance with the equation below following the Eurolab [27] technical report.
Precision of a research procedure in a laboratory is determined during validating the method or on the basis of measurements noted in control charts. Validation usually includes determining standard deviation of within-laboratory reproducibility srw or standard deviation of within -laboratory reproducibility sRw which is sometimes called intermediate precision. Bias is determined by means of standard deviation of measurement in relation to the reference value, e.g. while examining certified reference materials, using reference methods. The share of bias (b) in uncertainty of the measurement is determined with the help of mean deviation of measurements (Δ), uncertainty of reference material (uref ) and precision of measurements during examining that bias (s). The value of the expression s2 / n with a bigger number of measurements is low and can be omitted:
\n\t\t\tIn practice, different measurements result in different values of bias. In such a case the data may be combined and common assessment of a value of bias (uw) may be determined as a function of the measured value or, for typical data of matrices and levels, according to the formula below:
When certified reference materials are lacking (a frequent situation) and when no other analyses of bias have been performed in the laboratory (e.g. prior to applying the reference method) bias can be estimated on the basis of proficiency testing, PT.
A laboratory participating in PT may use the results of such tests in order to assess uncertainty of measurement for the testing method/procedure used. Similarly to determining uncertainty in within-laboratory experimental approach, the uncertainty of measurement (u) is equal to the root of the sum of squared values of standard deviation for within-laboratory reproducibility sRw and the bias (b), which can be calculated from the formulas 1,2 and 3.
With this approach two components of uncertainty are obtained from different sources. Precision is determined on the basis of the authors’ own validation data (within-laboratory reproducibility), from the range or on the basis of measuring control charts (in-house). The bias is determined on the basis of PT results. Estimating the bias on the basis of a single participation in PT may have a limited range and should be treated as preliminary. If the data from several PTs are available (a wider range of matrices and concentrations) the assessment of the bias may be referred to the complete measurement range.
The results of analysing uncertainty on the basis of experimental approaches using the results of the authors’ own results are presented below along with, for comparison, expanded uncertainties estimated with the help of Horwitz formula RSDR (%) = 2C-0.15.
Additive | Feed | bias (%) | u (%) | U = 2 ∙u (%) | U (%) * | |
Vitamin A | Feedingstuffs | 4.0 | 12.4 | 13.1 | 26.2 | 23.8 |
Premixes | 2.0 | 7.2 | 7.5 | 15.0 | 11.8 | |
Vitamin E | Feedingstuffs | 1.0 | 9.0 | 9.1 | 18.2 | 16.1 |
Premixes | 2.0 | 6.1 | 6.4 | 12.8 | 8.2 | |
Vitamin B1 | Feedingstuffs | 5.6 | 6.7 | 8.7 | 17.4 | 26.8 |
Premixes | 3.7 | 6.7 | 7.6 | 15.2 | 18.6 | |
Vitamin B2 | Feedingstuffs | 6.52 | 3.16 | 7.2 | 14.4 | 24.0 |
Premixes | 5.09 | 3.16 | 6.0 | 12.0 | 10.7 |
Results of uncertainty evaluation for some feed additives in compound feeds and premixes
The chapter presents a brief review of the methods used for determining feed additives by means of high proficiency liquid chromatography, HPLC. The authors presented their own research procedures and special attention was given to the preparation of samples for testing, extraction, extract purification, chromatographic separation and the basic elements of method validation and quality control.
Using HPLC for testing fat-soluble vitamins in feed materials, mixtures and premixes enabled us to replace colorimetric methods and to eliminate bias, such as the positive error of vitamin A determination related to the presence of carotenoids in the analyzed feed. The problem of low precision of examining certain vitamins, e.g. vitamin A, in feed mixtures is often unrelated to the method of determination, but rather to non-homogenous distribution of vitamin A in the feed related to its being secured against losing activity, due to protective coating. This problem may be solved by preparing the analytical weighed amount of sufficiently high mass and grinding the sample immediately prior to determination procedure to particles sized 1 mm.
Progress in the area of examining the content of water-soluble vitamins is also related to introducing the methods of liquid chromatography. The authors included their own procedures of analyzing vitamins B1 and B2, thiamine and riboflavin, with the use of HPLC methods and gave their characteristic parameters which meet the current requirements regarding the assessment of content and interpretation of results. These methods may be used especially to examine low content of thiamine and riboflavin, endogenic and added, in feed materials and mixtures.
HPLC methods have been widely used for testing cocciodiostats in feed preparations, premixes and mixtures. They contributed to improving the safety of using these additives, controlling concordance with manufacturer’s declaration and not exceeding the maximum content in feed mixtures, as well as controlling the withdrawal period. Without liquid chromatography with mass spectrometry (LCMS) it wouldn’t be possible to analyze effectively the remains of coccidiostats in the tissues and food products of animal origin. To reduce the risk of cross contamination in non-target feeds maximum content values for coccidiostats were determined recently at the level from 0.01 mg/kg (diclazuril) to 1.25 mg/kg (narasin, monensin), [29]. This created a need to develop some test methods adequate for the level of acceptable cross contamination and verifying them in interlaboratory tests. Future research will focus on checking the LCMS method for this particular purpose.
The official methods of separating and determining amino acids in feedingstuffs [13,8] are based mainly on ion exchange chromatography. However, in examining free amino acids (amino acids used as additives: lysine, methionine, threonine, tryptophan, valine, arginine and cysteine) HPLC methods are becoming increasingly more popular as they make the analyses shorter in time. In some cases a HCLP method is the only solution, e.g. while determining methionine hydroxy analog, verified in the authors’ own studies. The need to perform a large number of analyses in a shorter time determines the direction of future studies of amino acids in feedingstuffs and using ultra-performance liquid chromatography, UPLC, for this purpose.
In the testings of feed colorants the most frequently used means were spectrophotometric methods [13,8]. The diversity of feed products and the resulting changeability of matrix, as well as determining the maximum content of colorants in feed mixtures, were the reasons for searching for new methods of examining colorants, including HPLC. An example of such a method in reference to canthaxanthin and a procedure based on the authors’ own research is quoted in the present work. Future research in this respect will use the LCMS method to a higher degree as it enables detecting and determining several feed colorants in a single sample in view of cis-trans steroisomers.
The origins of sustainable development as a concept go back to the birth of environmental movements in the second half of the XIX century, which began with a romantic emphasis on nature and the importance for human beings to understand it. Its genesis dates back to the Industrial Revolution in England, as an attempt to awaken awareness about air pollution produced by the industries that had had rapid and sudden growth, in the context of a weak legal framework [1, 2, 3].
The discussion about the need of preserving the quality of natural resources joined previous debates on the durability of several raw materials, as well as on the degradation of the environment because of its use. That was the case when there was shortage of certain products and materials or the occurrence of local environmental problems associated with uncontrolled exploitation [4]. Some examples date back as far as Egyptian, Mesopotamian, Greek, and Roman civilizations, several thousand years before Christ, when evidence was collected of deforestation, salinization as well as soil loss linked to overexploitation of land resources in specific areas [4]. In more recent times, we can cite the shortage of wood caused by its excessive use in boat building, which severely threatened Europe in the XVIII century [4].
As a consequence of these episodes, the term “sustainable use” began to appear in forestry, as a way to guarantee the durability of wood exploitation, balancing the cutting of old trees with seedling planting [5].
Additionally, concerns over the excessive use of certain natural resources acquired other nuances in XVII and XVIII centuries, when different authors warned about the consequences of the overconsumption of natural resources, being The Tragedy of the Commons by Lloyd [6] and the Essay on the Principle of Population by Malthus [7] two of the most iconic.
In fact, this new perspective introduced a global vision into the debate, pointing out the existing contrast between a model based on unlimited growth of natural resources consumption and their inherent natural limitation in terms of volumes and times of recovery, not only at a local but also at a planetary scale. In this context, the XX century took place, with an exponential growth of studies that revealed the global climate crisis [8, 9, 10, 11, 12, 13].
Currently, despite the progress made defining a world framework around sustainable development and the assessment of environmental and social impacts, which includes the introduction of new goals [14] and standards (e.g. International Finance Corporation (IFC), performance standards), its implementation remains something difficult to achieve, simply because it is not compatible with the current economic model based on infinite growth and nature’s exploitation. The discussion remains focused on profit and return of investment.
To propose an alternate vision, we must reshape the current economic decisions through a wider lens of social, economic, environmental, and political sustainability. This includes a multi-stakeholder approach, where cooperation and synergy at different levels are two cornerstones. Private sector is a key stakeholder, since smart and forward-thinking entrepreneurs are an essential component for the change. They have a key role in identifying feasible ways to reduce environmental impacts and improve life conditions for local people, while working to generate economic and financial benefits [15]. This is possible when companies introduce the environment and local communities into the decision-making process [15].
The Dominican Republic hosts a series of characteristics that have fostered the development of alternatives and new narratives in local climate action, sustainable development, and social investment. The weakness of the Dominican national electric system, which does not guarantee neither universal access to electricity nor continuity of the service, promotes the emergence of alternative solutions which turn into innovative models for local sustainable development financed through social investment and sustained through a multi-stakeholder approach [16].
One of these solutions are community microhydropower systems, which, under the coordination of Guakía Ambiente and the Small Grants Programme (SGP/GEF/UNDP), transform local opportunities and social investment into integral development, based on the empowerment of women and men and the synergy among diverse stakeholders.
This essay discusses the increasing presence of social investment practices by the private sector in these initiatives [16]. By examining the main experiences and lessons learned obtained in more than 20 years of implementation, the document is intended to introduce the case study of microhydropower systems installed in the Dominican Republic and border region of Haiti, specifically focusing on the central role of social investment inside a multi-stakeholder scheme of intervention, as a catalyst of a sustainable model.
The results and conclusions are based on the analysis of technical documents and previous essays, which was complemented with the interview of members from Guakía Ambiente and the Small Grants Programme.
The concept of social investment has changed over the years and has been paired with many other concepts such as responsible investment, Environmental, Social, and Governance (ESG) investments, among others. Multilateral organizations and investment groups have developed their own definitions for social and responsible investment practices. However, most of them are aimed at reducing social conflicts, gaining reputation and maximizing profits in the long term. For example, the International Finance Corporation (IFC) developed the concept of community investing to “help companies gain a social license to operate, access land, reduce project and reputational risks, boost productivity, meet government requirements or global standards, and/or successfully compete for the next venture.” [17].
The same approach is used when speaking of ESG funds, where asset managers seek to keep the returns similar to traditional investments and do not necessarily aim at community development and sustainability at a local level [18]. Recovering the arguments from the previous chapter, this article seeks to understand social investment as something that directly impacts positively and empowers local populations, addresses climate challenges, and contributes to a long-term sustainability vision of a country.
The focus of the current document is not to discuss the concept of social investment, and it will use the definition launched by the United Nations Global Compact, which is broad enough to encompass different nuances regarding this complex and widely discussed concept [19].
Social investment was defined in 2010 by the United Nations (UN) as “the practice of making voluntary financial and nonfinancial contributions that demonstrably help local communities and broader societies to address their development priorities.” In order for an investment to be considered responsible and social, it must be purposeful, accountable, respectful, and ethical [20].
Additionally, this article focuses on the nonmonetary and nonprofit side of social investing. Private companies, foundations, and international cooperation make these investments for a wide variety of objectives. One of them is sustainability and climate action.
The urgency of actions to address the climate crisis needs creative solutions that do not necessarily give a monetary return to investors. However, there are several proxy indicators that can help to quantify the nonmonetary impact of social investments. For example, the number of hectares reforested and under community watershed management brings general benefits to air and water quality that are hard to express in monetary terms but largely contribute to the climate objectives of foundations and some private investors.
In this context, social investors have looked for initiatives that align with their sustainability objectives. In the Dominican Republic, a wide network of community microhydropower systems has developed thanks to these investments. In this section, the article will dissect this model of decentralized community power generation and its links with sustainability and social investment.
Located in the Caribbean, the Dominican Republic is a Small Island Developing State (SIDS) [21], which is highly vulnerable to the impacts of climate change [22], with high inequality and significant levels of poverty [23]. It is also one of the Latin American countries with the highest economic growth considering only the Gross Domestic Product (GDP) indicator [24].
In this context, in 1998, a pilot project of a microhydropower system was implemented seeking to address the lack of access to electricity of 70 families of the rural community of El Limón, located in the Southeastern portion of the Cordillera Central, the main massif of the country. From then on, the number of community microhydropower systems have grown exponentially in the country: 60 systems of this kind were built in the Dominican Republic and another one in the border region of Haiti, with a total combined installed capacity of 1.5 MW [25]. This ample network of micro-grids gives electricity and empowers more than 5000 households and 20,000 people in climate vulnerable mountainous areas with marginalized populations (Figure 1). In the process, over 70 square kilometers of land were restored and/or are being conserved in the direct and indirect area of influence of the microhydropower systems, while it is estimated that more than 25,000 tons of CO2 are absorbed and/or their emission avoided annually.
Basins where community microhydropower systems are working. [
Far beyond being a mere localized project, these systems promote an alternative to the energy transition in the Dominican Republic, where local stakeholders are active subjects in the fight against climate change. Decentralizing the power grid and giving the control of electricity flows to local populations promote empowerment and organization skills among men and women.
Moreover, the process of project implementation together with local communities is an example of how a just energy and people centered transition can be carried out. In fact, the starting point of the project is a specific and expressed need of a local group to solve their access to electricity. The solution to this need comes from a participatory process, where the community and its interests are at the center of the implementation, interacting synergically with other numerous external stakeholders.
Through a learning-by-doing process, community members and organizations train their abilities and acquire new skills. This way, the project turns into a capacity building experience, where people are free to learn and test their knowledge in a protected environment.
Nonmonetary benefits arising from these microhydropower systems are multiple. Most of them are hard to express in terms of monetary value, such as empowerment or capacity building. This difficulty is present even in costs internationally linked to policy, such as the social carbon price, which can range from lows as 25 to highs as 800 dollars per ton [26, 27].
Due to the complexity of interlinked environmental, political, and social systems, quantifying community and environmental benefits is probably not the best strategy to assess the positive impacts and assign a value of return to the social investments in these initiatives. For instance, community microhydropower systems are based on community capitals and the promotion of sustainable livelihoods [28, 29, 30, 31, 32, 33], instead of the promotion of mere financial capital gains or profit.
Following this idea, we can analyze the benefits through quantitative data of environmental and social improvements, as well as qualitative indicators. The scope of this article is not to generate a system of qualitative and quantitative assessment of the impact of community hydropower systems. However, this section will disclose several of the social and environmental benefits that can be used to further build a robust assessment system.
One of the most general benefits from the social investment in community microhydropower is the advancing toward the accomplishment of climate goals and more specifically of working toward a just energy transition. It is also implemented under a perspective of Nature-based Solutions (NbS) that brings together efforts of numerous stakeholders, who make synergy to reach a common objective (Figure 2).
Sustainable model from the community microhydropower systems.
Community microhydro systems contribute to carry out interventions that “protect, sustainably manage, and restore natural and modified ecosystems that address societal challenges effectively and adaptively, simultaneously providing human well-being and biodiversity benefits,” aligned to the definition that the International Union for Conservation of Nature (IUCN) provides for NbS [34].
The following Table 1 summarizes the categories and proposes basic initial indicators where community microhydropower systems impact, which can be used as nonmonetary success indicators in terms of social investment:
Category | Indicators |
---|---|
Decentralized generation from a renewable energy source |
|
Energy efficiency |
|
Productive use of energy |
|
Forest conservation and watershed management |
|
Capacity building |
|
Local empowerment, gender equality, and social inclusion |
|
Socio environmental and economic benefits |
|
Climate change mitigation and adaptation |
|
Basic indicators to measure the nonmonetary success of social investment.
Using merely traditional financial indicators, community microhydropower systems might not be the best financial option. However, if we use the previous indicators and, at the same time, we are able to quantify the climate and social benefits, they might as well be profitable. For example, the costs associated with a typical 20-kW system are $300,000, of which around 75% is not refundable if you consider only economic indicators. This means that, if we want to apply a social investment framework, we must build new nonmonetary ways of quantifying returns of investment, which includes the previous indicators.
In this process, the role of the private sector and its social investments in the community microhydropower systems has been progressively growing in the country and becoming a significant component of the implementation of the projects. Likewise, this interaction among civil society organizations, community members, government officials, and private entities has strengthened the Corporate Social Responsibility policy of several national companies and is becoming central to their social and business agenda.
Among them, the action of the Popular Group (Grupo Popular) stands out. It is one of the main financial groups of the Dominican Republic, which has been increasing its action in these initiatives through the Popular Foundation (Fundación Popular), the entity responsible for its social investment agenda, coming across an effective scheme to contribute to integral interventions, overcoming the limitations of specific projects outside of a strategic framework.
Each community microhydro project is financed through concessional investments, basically different types of nonrefundable investments such as seed capital, grants, and donations. Nevertheless, this has not stopped private investors, such as Grupo Popular, to step in and assign a social investment budget to fuel the energy transition and community development. The framework private firms are using to assess the impact of their investments that are not monetary and rely on the idea of community development and sustainability.
To this day, we can estimate that the total investment made in the implementation of microhydro projects under this scheme exceeds 17 million dollars. The social investment from the private sector is around 5% of it, with a significant increase in its participation during the last 10 years. There is still plenty of opportunity for social investors to increase their participation in these projects.
The participation of the private sector in these projects is an alternative to reduce the risks associated with their implementation. Moreover, multi-stakeholder approaches can fuel a virtuous circle of local development based on social investment and sustainability.
Access to basic services, like electricity, is an essential assumption for development. Nevertheless, sustainability is rather strictly linked to social and environmental processes and dynamics that can be promoted among human groups and their territories. According to this perspective, each project can lighten a path to implement coordinated multi-stakeholder processes, aimed to reach wider sustainability goals.
From this point of view, empowerment of people and local organized groups should become a guiding principle for each of the stakeholders who take part in the process, from private entities to government officials. There should be a common effort to strengthen personal and community self-esteem: during the process, people, individually and as a part of a human group, should internalize that they can reach previously planned goals.
These interventions contribute to educating people to freedom, providing them the opportunity to acquire knowledge and instruments that allow them to make decisions and contribute to the development of the social group they belong to. Confidence, especially inside the community, is a key issue: the intervention should help to overcome the common situation where people show more confidence toward outsiders than toward their peers.
Another important triggering factor is the promotion of solidarity among and within stakeholders. The principle according to which maximizing individual benefit would maximize the collective one has revealed all its limitations, especially when it is applied without a vision of social well-being and sustainability, transcending the idea of only seeking economic benefit. The most successful and sustainable societies are those who adopt synergic strategies and invest in their own development. This includes mechanisms of mutual support, which allow people and communities to overcome individual and collective challenges, generating a sense of belonging to the social group and the environment where they live. Solidarity is a key element to increase the positive impacts of social investments, since it promotes a spiral of continuous improvement.
From this point of view, this model of intervention contributes to improving cost-effectiveness, promoting appropriate distribution of the tasks and available resources, avoiding duplicity, and facilitating the elaboration and implementation of middle- and long-term plans.
According to the experience gained in multiple projects, the development and social investment models must reintroduce the concept of sustainability along with ethics and nonmonetary objectives and indicators. Community development is a complex and slow process, where empowerment is a key to achieving goals.
Social investment in the country has fulfilled a role of filling in the gap of financing projects on a nonrefundable basis. Most of the projects need nonrefundable investments, since the return of investment, measured only in financial terms, would render most of the projects unfeasible for a traditional financial model. However, if there was a way to incorporate social and environmental positive impacts and sustainable development in the framework of financing decisions of other institutions, it would be possible to expand the impact of social investments.
There is also a need to go further in the models of community microhydropower systems in order to create mechanisms to provide monetary returns in order to diversify the source of social investments. It is also possible to take advantage of the existing network of community microhydropower systems and their current savings networks.
There is the example of the community of Angostura, where the first project was implemented through a combination of social investment, donations by international cooperation, and seed capital. Afterward, the local economy was reshaped, and an ecotourism project was developed, as well as a simple but complete self-managed system of monthly payments to a community fund. With these resources, Angostura was able to participate in covering the costs of an expansion of the microhydropower generation, providing more than 10,000 dollars from its community fund.
The use and abuse of natural resources has become an increasingly critical issue to the human and nonhuman population, impairing the human group’s capacity to satisfy its own needs. The multidimensional climate crisis is being fueled not only by the burning of fossil fuels but also by the depletion of ecosystems. This has caused an exponentially growing pressure over the environment through unsustainable investments and projects based on a development model that is incompatible with the natural limits of the resources.
In this context, the expected solution should go through a paradigm shift based on empowerment of people and communities, strengthening of solidarity across sectors and stakeholders and finding creative solutions to social investment opportunities. Communities should acquire means and skills to participate freely, actively, and significatively in decision-making processes of development, at any level.
The evidence coming from the experience of community microhydropower systems points out that the reintroduction of ethics in decision-making is fundamental to reach sustainability. In fact, even though the first step is the development of strategies and policies based on a model that is compatible with the recovery and reposition of natural resources, the effectiveness and durability of its implementation are strictly linked to the commitment of the local population in doing so and the cooperation with other stakeholders, far beyond the regime of consequences that has been established.
Based on well-known technology, these initiatives have become a reference of just energy transition experiences, climate action, and integral watershed management, where local communities, interacting with numerous stakeholders in fair conditions, assume the role of guiding their own development, which includes the monitoring and caring of the natural resources in their territories.
Up until now the social investment by the private sector in these projects has increased, but in absolute terms it is still limited. By increasing the participation of the private sector in these climate solutions, based on synergies among stakeholders, such as government agencies, communities, and international cooperation, there is an opportunity for replicating and upscaling these initiatives. As well, risks associated with funding, timings, governance, government changes, among others, can be significantly reduced.
Nonmonetary assessments such as the ones proposed in this paper are necessary to improve the analysis of the return of the social investments, overcoming the present limits and biases of financial assessments. It is also essential to build a framework around social and environmental indicators in order to build a paradigm shift in climate investment. In this sense, Guakía Ambiente, the Small Grants Programme, and other stakeholders are working to face this challenge.
Guakía Ambiente is a Dominican-based nonprofit organization which promotes the development and empowerment of local communities through renewable energy and nature based solutions. More information at: www.guakiambiente.org.
The Small Grants Program (PPS-SGP) is an initiative of the Global Environment Facility (GEF) aimed at supporting civil society organizations with non-reimbursable funds and technical-administrative support in the development of community actions in favor of the global environment and generates well-being for people at the local level. More information at: www.ppsdom.org
This work was supported by the Interamerican Foundation [Project DR-347 “Acceso a servicio de electricidad en comunidades rurales de la República Dominicana mediante instalación de cuatro sistemas micro hidroeléctricos comunitarios”].
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Aalborg University has Two Satellite Campuses, one in Copenhagen (Aalborg University Copenhagen) and the other in Esbjerg (Aalborg University Esbjerg).\n· He is a member of prestigious IEEE (Institute of Electrical and Electronics Engineers), and IAENG (International Association of Engineers) organizations. \n· He is the chief Editor of the Journal of Software Engineering.\n· He is the member of the Editorial Board of International Journal of Computer Science and Software Technology (IJCSST) and International Journal of Computer Engineering and Information Technology. \n· He is also the Editor of Communication in Computer and Information Science CCIS-20 by Springer.\n· Reviewer For Many Conferences\nHe is the lead person in making collaboration agreements between Aalborg University and many universities of Pakistan, for which the MOU’s (Memorandum of Understanding) have been signed.\nProfessor Akbar is working in Academia since 1990, he started his career as a Lab demonstrator/TA at the University of Sussex. After finishing his P. hD degree in 1992, he served in the Industry as a Scientific Officer and continued his academic career as a visiting scholar for a number of educational institutions. In 1996 he joined National University of Science & Technology Pakistan (NUST) as an Associate Professor; NUST is one of the top few universities in Pakistan. In 1999 he joined an International Company Lineo Inc, Canada as Manager Compiler Group, where he headed the group for developing Compiler Tool Chain and Porting of Operating Systems for the BLACKfin processor. The processor development was a joint venture by Intel and Analog Devices. In 2002 Lineo Inc., was taken over by another company, so he joined Aalborg University Denmark as an Assistant Professor.\nProfessor Akbar has truly a multi-disciplined career and he continued his legacy and making progress in many areas of his interests both in teaching and research. 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While manufacturing nonwovens, some conventional textile operations, such as carding, drawing, roving, spinning, weaving or knitting, are partially or completely eliminated. For this reason the choice of fiber is very important for nonwoven manufacturers. The commonly used fibers include natural fibers (cotton, jute, flax, wool), synthetic fibers (polyester (PES), polypropylene (PP), polyamide, rayon), special fibers (glass, carbon, nanofiber, bi-component, superabsorbent fibers). Raw materials have not only delivered significant product improvements but also benefited people using these products by providing hygiene and comfort.",book:{id:"5062",slug:"non-woven-fabrics",title:"Non-woven Fabrics",fullTitle:"Non-woven Fabrics"},signatures:"Nazan Avcioglu Kalebek and Osman Babaarslan",authors:[{id:"119775",title:"Prof.",name:"Osman",middleName:null,surname:"Babaarslan",slug:"osman-babaarslan",fullName:"Osman Babaarslan"},{id:"175829",title:"Dr.",name:"Nazan",middleName:null,surname:"Kalebek",slug:"nazan-kalebek",fullName:"Nazan Kalebek"}]},{id:"41409",title:"Surface Modification Methods for Improving the Dyeability of Textile Fabrics",slug:"surface-modification-methods-for-improving-the-dyeability-of-textile-fabrics",totalDownloads:7063,totalCrossrefCites:13,totalDimensionsCites:36,abstract:null,book:{id:"3137",slug:"eco-friendly-textile-dyeing-and-finishing",title:"Eco-Friendly Textile Dyeing and Finishing",fullTitle:"Eco-Friendly Textile Dyeing and Finishing"},signatures:"Sheila Shahidi, Jakub Wiener and Mahmood Ghoranneviss",authors:[{id:"58854",title:"Dr.",name:null,middleName:null,surname:"Shahidi",slug:"shahidi",fullName:"Shahidi"}]},{id:"55424",title:"Textile Reinforced Structural Composites for Advanced Applications",slug:"textile-reinforced-structural-composites-for-advanced-applications",totalDownloads:3876,totalCrossrefCites:9,totalDimensionsCites:16,abstract:"Textile-reinforced composites are increasingly used in various industries such as aerospace, construction, automotive, medicine, and sports due to their distinctive advantages over traditional materials such as metals and ceramics. 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He previously worked as a post-doctoral fellow at the Ben-Gurion University of Negev, Israel; University of the Free State, South Africa; and Central University of Technology Bloemfontein, South Africa. He obtained his Ph.D. in Organic Chemistry from Nagaoka University of Technology, Japan. He has published more than seventy-four journal articles and attended several national and international conferences as speaker and chair. Dr. Kendrekar has received many international awards. He has several funded projects, namely, anti-malaria drug development, MRSA, and SARS-CoV-2 activity of curcumin and its formulations. He has filed four patents in collaboration with the University of Central Lancashire and Mayo Clinic Infectious Diseases. His present research includes organic synthesis, drug discovery and development, biochemistry, nanoscience, and nanotechnology.",institutionString:"Visiting Scientist at Lipid Nanostructures Laboratory, Centre for Smart Materials, School of Natural Sciences, University of Central Lancashire",institution:null},{id:"428125",title:"Dr.",name:"Vinayak",middleName:null,surname:"Adimule",slug:"vinayak-adimule",fullName:"Vinayak Adimule",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/428125/images/system/428125.jpg",biography:"Dr. Vinayak Adimule, MSc, Ph.D., is a professor and dean of R&D, Angadi Institute of Technology and Management, India. He has 15 years of research experience as a senior research scientist and associate research scientist in R&D organizations. He has published more than fifty research articles as well as several book chapters. He has two Indian patents and two international patents to his credit. Dr. Adimule has attended, chaired, and presented papers at national and international conferences. He is a guest editor for Topics in Catalysis and other journals. He is also an editorial board member, life member, and associate member for many international societies and research institutions. His research interests include nanoelectronics, material chemistry, artificial intelligence, sensors and actuators, bio-nanomaterials, and medicinal chemistry.",institutionString:"Angadi Institute of Technology and Management",institution:null},{id:"284317",title:"Prof.",name:"Kantharaju",middleName:null,surname:"Kamanna",slug:"kantharaju-kamanna",fullName:"Kantharaju Kamanna",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/284317/images/21050_n.jpg",biography:"Prof. K. Kantharaju has received Bachelor of science (PCM), master of science (Organic Chemistry) and Doctor of Philosophy in Chemistry from Bangalore University. He worked as a Executive Research & Development @ Cadila Pharmaceuticals Ltd, Ahmedabad. He received DBT-postdoc fellow @ Molecular Biophysics Unit, Indian Institute of Science, Bangalore under the supervision of Prof. P. Balaram, later he moved to NIH-postdoc researcher at Drexel University College of Medicine, Philadelphia, USA, after his return from postdoc joined NITK-Surthakal as a Adhoc faculty at department of chemistry. Since from August 2013 working as a Associate Professor, and in 2016 promoted to Profeesor in the School of Basic Sciences: Department of Chemistry and having 20 years of teaching and research experiences.",institutionString:null,institution:{name:"Rani Channamma University, Belagavi",country:{name:"India"}}},{id:"158492",title:"Prof.",name:"Yusuf",middleName:null,surname:"Tutar",slug:"yusuf-tutar",fullName:"Yusuf Tutar",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/158492/images/system/158492.jpeg",biography:"Prof. Dr. Yusuf Tutar conducts his research at the Hamidiye Faculty of Pharmacy, Department of Basic Pharmaceutical Sciences, Division of Biochemistry, University of Health Sciences, Turkey. He is also a faculty member in the Molecular Oncology Program. He obtained his MSc and Ph.D. at Oregon State University and Texas Tech University, respectively. He pursued his postdoctoral studies at Rutgers University Medical School and the National Institutes of Health (NIH/NIDDK), USA. His research focuses on biochemistry, biophysics, genetics, molecular biology, and molecular medicine with specialization in the fields of drug design, protein structure-function, protein folding, prions, microRNA, pseudogenes, molecular cancer, epigenetics, metabolites, proteomics, genomics, protein expression, and characterization by spectroscopic and calorimetric methods.",institutionString:"University of Health Sciences",institution:null},{id:"180528",title:"Dr.",name:"Hiroyuki",middleName:null,surname:"Kagechika",slug:"hiroyuki-kagechika",fullName:"Hiroyuki Kagechika",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/180528/images/system/180528.jpg",biography:"Hiroyuki Kagechika received his bachelor’s degree and Ph.D. in Pharmaceutical Sciences from the University of Tokyo, Japan, where he served as an associate professor until 2004. He is currently a professor at the Institute of Biomaterials and Bioengineering (IBB), Tokyo Medical and Dental University (TMDU). From 2010 to 2012, he was the dean of the Graduate School of Biomedical Science. Since 2012, he has served as the vice dean of the Graduate School of Medical and Dental Sciences. He has been the director of the IBB since 2020. Dr. Kagechika’s major research interests are the medicinal chemistry of retinoids, vitamins D/K, and nuclear receptors. He has developed various compounds including a drug for acute promyelocytic leukemia.",institutionString:"Tokyo Medical and Dental University",institution:{name:"Tokyo Medical and Dental University",country:{name:"Japan"}}},{id:"94311",title:"Prof.",name:"Martins",middleName:"Ochubiojo",surname:"Ochubiojo Emeje",slug:"martins-ochubiojo-emeje",fullName:"Martins Ochubiojo Emeje",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/94311/images/system/94311.jpeg",biography:"Martins Emeje obtained a BPharm with distinction from Ahmadu Bello University, Nigeria, and an MPharm and Ph.D. from the University of Nigeria (UNN), where he received the best Ph.D. award and was enlisted as UNN’s “Face of Research.” He established the first nanomedicine center in Nigeria and was the pioneer head of the intellectual property and technology transfer as well as the technology innovation and support center. Prof. Emeje’s several international fellowships include the prestigious Raman fellowship. He has published more than 150 articles and patents. He is also the head of R&D at NIPRD and holds a visiting professor position at Nnamdi Azikiwe University, Nigeria. He has a postgraduate certificate in Project Management from Walden University, Minnesota, as well as a professional teaching certificate and a World Bank certification in Public Procurement. Prof. Emeje was a national chairman of academic pharmacists in Nigeria and the 2021 winner of the May & Baker Nigeria Plc–sponsored prize for professional service in research and innovation.",institutionString:"National Institute for Pharmaceutical Research and Development",institution:{name:"National Institute for Pharmaceutical Research and Development",country:{name:"Nigeria"}}},{id:"436430",title:"Associate Prof.",name:"Mesut",middleName:null,surname:"Işık",slug:"mesut-isik",fullName:"Mesut Işık",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/436430/images/19686_n.jpg",biography:null,institutionString:null,institution:{name:"Bilecik University",country:{name:"Turkey"}}},{id:"268659",title:"Ms.",name:"Xianquan",middleName:null,surname:"Zhan",slug:"xianquan-zhan",fullName:"Xianquan Zhan",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/268659/images/8143_n.jpg",biography:"Dr. Zhan received his undergraduate and graduate training in the fields of preventive medicine and epidemiology and statistics at the West China University of Medical Sciences in China during 1989 to 1999. He received his post-doctoral training in oncology and cancer proteomics for two years at the Cancer Research Institute of Human Medical University in China. In 2001, he went to the University of Tennessee Health Science Center (UTHSC) in USA, where he was a post-doctoral researcher and focused on mass spectrometry and cancer proteomics. Then, he was appointed as an Assistant Professor of Neurology, UTHSC in 2005. He moved to the Cleveland Clinic in USA as a Project Scientist/Staff in 2006 where he focused on the studies of eye disease proteomics and biomarkers. He returned to UTHSC as an Assistant Professor of Neurology in the end of 2007, engaging in proteomics and biomarker studies of lung diseases and brain tumors, and initiating the studies of predictive, preventive, and personalized medicine (PPPM) in cancer. In 2010, he was promoted to Associate Professor of Neurology, UTHSC. Currently, he is a Professor at Xiangya Hospital of Central South University in China, Fellow of Royal Society of Medicine (FRSM), the European EPMA National Representative in China, Regular Member of American Association for the Advancement of Science (AAAS), European Cooperation of Science and Technology (e-COST) grant evaluator, Associate Editors of BMC Genomics, BMC Medical Genomics, EPMA Journal, and Frontiers in Endocrinology, Executive Editor-in-Chief of Med One. He has\npublished 116 peer-reviewed research articles, 16 book chapters, 2 books, and 2 US patents. His current main research interest focuses on the studies of cancer proteomics and biomarkers, and the use of modern omics techniques and systems biology for PPPM in cancer, and on the development and use of 2DE-LC/MS for the large-scale study of human proteoforms.",institutionString:null,institution:{name:"Xiangya Hospital Central South University",country:{name:"China"}}},{id:"40482",title:null,name:"Rizwan",middleName:null,surname:"Ahmad",slug:"rizwan-ahmad",fullName:"Rizwan Ahmad",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/40482/images/system/40482.jpeg",biography:"Dr. Rizwan Ahmad is a University Professor and Coordinator, Quality and Development, College of Medicine, Imam Abdulrahman bin Faisal University, Saudi Arabia. Previously, he was Associate Professor of Human Function, Oman Medical College, Oman, and SBS University, Dehradun. Dr. Ahmad completed his education at Aligarh Muslim University, Aligarh. He has published several articles in peer-reviewed journals, chapters, and edited books. His area of specialization is free radical biochemistry and autoimmune diseases.",institutionString:"Imam Abdulrahman Bin Faisal University",institution:{name:"Imam Abdulrahman Bin Faisal University",country:{name:"Saudi Arabia"}}},{id:"41865",title:"Prof.",name:"Farid A.",middleName:null,surname:"Badria",slug:"farid-a.-badria",fullName:"Farid A. Badria",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/41865/images/system/41865.jpg",biography:"Farid A. Badria, Ph.D., is the recipient of several awards, including The World Academy of Sciences (TWAS) Prize for Public Understanding of Science; the World Intellectual Property Organization (WIPO) Gold Medal for best invention; Outstanding Arab Scholar, Kuwait; and the Khwarizmi International Award, Iran. He has 250 publications, 12 books, 20 patents, and several marketed pharmaceutical products to his credit. He continues to lead research projects on developing new therapies for liver, skin disorders, and cancer. Dr. Badria was listed among the world’s top 2% of scientists in medicinal and biomolecular chemistry in 2019 and 2020. He is a member of the Arab Development Fund, Kuwait; International Cell Research Organization–United Nations Educational, Scientific and Cultural Organization (ICRO–UNESCO), Chile; and UNESCO Biotechnology France",institutionString:"Mansoura University",institution:{name:"Mansoura University",country:{name:"Egypt"}}},{id:"329385",title:"Dr.",name:"Rajesh K.",middleName:"Kumar",surname:"Singh",slug:"rajesh-k.-singh",fullName:"Rajesh K. Singh",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/329385/images/system/329385.png",biography:"Dr. Singh received a BPharm (2003) and MPharm (2005) from Panjab University, Chandigarh, India, and a Ph.D. (2013) from Punjab Technical University (PTU), Jalandhar, India. He has more than sixteen years of teaching experience and has supervised numerous postgraduate and Ph.D. students. He has to his credit more than seventy papers in SCI- and SCOPUS-indexed journals, fifty-five conference proceedings, four books, six Best Paper Awards, and five projects from different government agencies. He is currently an editorial board member of eight international journals and a reviewer for more than fifty scientific journals. He received Top Reviewer and Excellent Peer Reviewer Awards from Publons in 2016 and 2017, respectively. He is also on the panel of The International Reviewer for reviewing research proposals for grants from the Royal Society. He also serves as a Publons Academy mentor and Bentham brand ambassador.",institutionString:"Punjab Technical University",institution:{name:"Punjab Technical University",country:{name:"India"}}},{id:"142388",title:"Dr.",name:"Thiago",middleName:"Gomes",surname:"Gomes Heck",slug:"thiago-gomes-heck",fullName:"Thiago Gomes Heck",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/142388/images/7259_n.jpg",biography:null,institutionString:null,institution:{name:"Universidade Regional do Noroeste do Estado do Rio Grande do Sul",country:{name:"Brazil"}}},{id:"336273",title:"Assistant Prof.",name:"Janja",middleName:null,surname:"Zupan",slug:"janja-zupan",fullName:"Janja Zupan",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/336273/images/14853_n.jpeg",biography:"Janja Zupan graduated in 2005 at the Department of Clinical Biochemistry (superviser prof. dr. Janja Marc) in the field of genetics of osteoporosis. Since November 2009 she is working as a Teaching Assistant at the Faculty of Pharmacy, Department of Clinical Biochemistry. In 2011 she completed part of her research and PhD work at Institute of Genetics and Molecular Medicine, University of Edinburgh. She finished her PhD entitled The influence of the proinflammatory cytokines on the RANK/RANKL/OPG in bone tissue of osteoporotic and osteoarthritic patients in 2012. From 2014-2016 she worked at the Institute of Biomedical Sciences, University of Aberdeen as a postdoctoral research fellow on UK Arthritis research project where she gained knowledge in mesenchymal stem cells and regenerative medicine. She returned back to University of Ljubljana, Faculty of Pharmacy in 2016. She is currently leading project entitled Mesenchymal stem cells-the keepers of tissue endogenous regenerative capacity facing up to aging of the musculoskeletal system funded by Slovenian Research Agency.",institutionString:null,institution:{name:"University of Ljubljana",country:{name:"Slovenia"}}},{id:"357453",title:"Dr.",name:"Radheshyam",middleName:null,surname:"Maurya",slug:"radheshyam-maurya",fullName:"Radheshyam Maurya",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/357453/images/16535_n.jpg",biography:null,institutionString:null,institution:{name:"University of Hyderabad",country:{name:"India"}}},{id:"418340",title:"Dr.",name:"Jyotirmoi",middleName:null,surname:"Aich",slug:"jyotirmoi-aich",fullName:"Jyotirmoi Aich",position:null,profilePictureURL:"https://s3.us-east-1.amazonaws.com/intech-files/0033Y000038Ugi5QAC/Profile_Picture_2022-04-15T07:48:28.png",biography:"Biotechnologist with 15 years of research including 6 years of teaching experience. Demonstrated record of scientific achievements through consistent publication record (H index = 13, with 874 citations) in high impact journals such as Nature Communications, Oncotarget, Annals of Oncology, PNAS, and AJRCCM, etc. Strong research professional with a post-doctorate from ACTREC where I gained experimental oncology experience in clinical settings and a doctorate from IGIB where I gained expertise in asthma pathophysiology. A well-trained biotechnologist with diverse experience on the bench across different research themes ranging from asthma to cancer and other infectious diseases. An individual with a strong commitment and innovative mindset. Have the ability to work on diverse projects such as regenerative and molecular medicine with an overall mindset of improving healthcare.",institutionString:"DY Patil Deemed to Be University",institution:null},{id:"349288",title:"Prof.",name:"Soumya",middleName:null,surname:"Basu",slug:"soumya-basu",fullName:"Soumya Basu",position:null,profilePictureURL:"https://s3.us-east-1.amazonaws.com/intech-files/0033Y000035QxIDQA0/Profile_Picture_2022-04-15T07:47:01.jpg",biography:"Soumya Basu, Ph.D., is currently working as an Associate Professor at Dr. D. Y. Patil Biotechnology and Bioinformatics Institute, Dr. D. Y. Patil Vidyapeeth, Pune, Maharashtra, India. With 16+ years of trans-disciplinary research experience in Drug Design, development, and pre-clinical validation; 20+ research article publications in journals of repute, 9+ years of teaching experience, trained with cross-disciplinary education, Dr. Basu is a life-long learner and always thrives for new challenges.\r\nHer research area is the design and synthesis of small molecule partial agonists of PPAR-γ in lung cancer. She is also using artificial intelligence and deep learning methods to understand the exosomal miRNA’s role in cancer metastasis. Dr. Basu is the recipient of many awards including the Early Career Research Award from the Department of Science and Technology, Govt. of India. She is a reviewer of many journals like Molecular Biology Reports, Frontiers in Oncology, RSC Advances, PLOS ONE, Journal of Biomolecular Structure & Dynamics, Journal of Molecular Graphics and Modelling, etc. She has edited and authored/co-authored 21 journal papers, 3 book chapters, and 15 abstracts. She is a Board of Studies member at her university. She is a life member of 'The Cytometry Society”-in India and 'All India Cell Biology Society”- in India.",institutionString:"Dr. D.Y. Patil Vidyapeeth, Pune",institution:{name:"Dr. D.Y. Patil Vidyapeeth, Pune",country:{name:"India"}}},{id:"354817",title:"Dr.",name:"Anubhab",middleName:null,surname:"Mukherjee",slug:"anubhab-mukherjee",fullName:"Anubhab Mukherjee",position:null,profilePictureURL:"https://intech-files.s3.amazonaws.com/0033Y0000365PbRQAU/ProfilePicture%202022-04-15%2005%3A11%3A18.480",biography:"A former member of Laboratory of Nanomedicine, Brigham and Women’s Hospital, Harvard University, Boston, USA, Dr. Anubhab Mukherjee is an ardent votary of science who strives to make an impact in the lives of those afflicted with cancer and other chronic/acute ailments. He completed his Ph.D. from CSIR-Indian Institute of Chemical Technology, Hyderabad, India, having been skilled with RNAi, liposomal drug delivery, preclinical cell and animal studies. He pursued post-doctoral research at College of Pharmacy, Health Science Center, Texas A & M University and was involved in another postdoctoral research at Department of Translational Neurosciences and Neurotherapeutics, John Wayne Cancer Institute, Santa Monica, California. In 2015, he worked in Harvard-MIT Health Sciences & Technology as a visiting scientist. He has substantial experience in nanotechnology-based formulation development and successfully served various Indian organizations to develop pharmaceuticals and nutraceutical products. He is an inventor in many US patents and an author in many peer-reviewed articles, book chapters and books published in various media of international repute. Dr. Mukherjee is currently serving as Principal Scientist, R&D at Esperer Onco Nutrition (EON) Pvt. Ltd. and heads the Hyderabad R&D center of the organization.",institutionString:"Esperer Onco Nutrition Pvt Ltd.",institution:null},{id:"319365",title:"Assistant Prof.",name:"Manash K.",middleName:null,surname:"Paul",slug:"manash-k.-paul",fullName:"Manash K. Paul",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/319365/images/system/319365.png",biography:"Manash K. Paul is a Principal Investigator and Scientist at the University of California Los Angeles. He has contributed significantly to the fields of stem cell biology, regenerative medicine, and lung cancer. His research focuses on various signaling processes involved in maintaining stem cell homeostasis during the injury-repair process, deciphering lung stem cell niche, pulmonary disease modeling, immuno-oncology, and drug discovery. He is currently investigating the role of extracellular vesicles in premalignant lung cell migration and detecting the metastatic phenotype of lung cancer via machine-learning-based analyses of exosomal signatures. Dr. Paul has published in more than fifty peer-reviewed international journals and is highly cited. 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Graduated in Pharmacy, specialization in Cosmetology and Cosmeceuticals applied to aesthetics, specialization in Aesthetic and Cosmetic Health, and a doctorate in Pharmaceutical Nanotechnology. Teaching experience in Pharmacy and Aesthetics and Cosmetics courses. She works mainly on the following subjects: nanotechnology, cosmetology, pharmaceutical technology, aesthetics.",institutionString:"Universidade Federal de Juiz de Fora",institution:{name:"Universidade Federal de Juiz de Fora",country:{name:"Brazil"}}},{id:"219081",title:"Dr.",name:"Abdulsamed",middleName:null,surname:"Kükürt",slug:"abdulsamed-kukurt",fullName:"Abdulsamed Kükürt",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/219081/images/system/219081.png",biography:"Dr. Kükürt graduated from Uludağ University in Turkey. He started his academic career as a Research Assistant in the Department of Biochemistry at Kafkas University. In 2019, he completed his Ph.D. program in the Department of Biochemistry at the Institute of Health Sciences. He is currently working at the Department of Biochemistry, Kafkas University. He has 27 published research articles in academic journals, 11 book chapters, and 37 papers. He took part in 10 academic projects. He served as a reviewer for many articles. He still serves as a member of the review board in many academic journals. 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He has 60 articles published in scientific journals and 20 poster presentations in scientific congresses. His research interests include physiology, endocrine system, cancer, diabetes, cardiovascular system diseases, and isolated organ bath system studies.",institutionString:"Kafkas University",institution:{name:"Kafkas University",country:{name:"Turkey"}}},{id:"418963",title:"Dr.",name:"Augustine Ododo",middleName:"Augustine",surname:"Osagie",slug:"augustine-ododo-osagie",fullName:"Augustine Ododo Osagie",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/418963/images/16900_n.jpg",biography:"Born into the family of Osagie, a prince of the Benin Kingdom. I am currently an academic in the Department of Medical Biochemistry, University of Benin. 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She is a fellow member of the Royal Society of Chemistry UK and the American Chemical Society of the United States.",institutionString:"King Saud University",institution:{name:"King Saud University",country:{name:"Saudi Arabia"}}},{id:"49848",title:"Dr.",name:"Wen-Long",middleName:null,surname:"Hu",slug:"wen-long-hu",fullName:"Wen-Long Hu",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/49848/images/system/49848.jpg",biography:"Wen-Long Hu is Chief of the Division of Acupuncture, Department of Chinese Medicine at Kaohsiung Chang Gung Memorial Hospital, as well as an adjunct associate professor at Fooyin University and Kaohsiung Medical University. Wen-Long is President of Taiwan Traditional Chinese Medicine Medical Association. He has 28 years of experience in clinical practice in laser acupuncture therapy and 34 years in acupuncture. He is an invited speaker for lectures and workshops in laser acupuncture at many symposiums held by medical associations. He owns the patent for herbal preparation and producing, and for the supercritical fluid-treated needle. Dr. Hu has published three books, 12 book chapters, and more than 30 papers in reputed journals, besides serving as an editorial board member of repute.",institutionString:"Kaohsiung Chang Gung Memorial Hospital",institution:{name:"Kaohsiung Chang Gung Memorial Hospital",country:{name:"Taiwan"}}},{id:"298472",title:"Prof.",name:"Andrey V.",middleName:null,surname:"Grechko",slug:"andrey-v.-grechko",fullName:"Andrey V. Grechko",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/298472/images/system/298472.png",biography:"Andrey Vyacheslavovich Grechko, Ph.D., Professor, is a Corresponding Member of the Russian Academy of Sciences. He graduated from the Semashko Moscow Medical Institute (Semashko National Research Institute of Public Health) with a degree in Medicine (1998), the Clinical Department of Dermatovenerology (2000), and received a second higher education in Psychology (2009). Professor A.V. Grechko held the position of Сhief Physician of the Central Clinical Hospital in Moscow. He worked as a professor at the faculty and was engaged in scientific research at the Medical University. Starting in 2013, he has been the initiator of the creation of the Federal Scientific and Clinical Center for Intensive Care and Rehabilitology, Moscow, Russian Federation, where he also serves as Director since 2015. He has many years of experience in research and teaching in various fields of medicine, is an author/co-author of more than 200 scientific publications, 13 patents, 15 medical books/chapters, including Chapter in Book «Metabolomics», IntechOpen, 2020 «Metabolomic Discovery of Microbiota Dysfunction as the Cause of Pathology».",institutionString:"Federal Research and Clinical Center of Intensive Care Medicine and Rehabilitology",institution:null},{id:"199461",title:"Prof.",name:"Natalia V.",middleName:null,surname:"Beloborodova",slug:"natalia-v.-beloborodova",fullName:"Natalia V. Beloborodova",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/199461/images/system/199461.jpg",biography:'Natalia Vladimirovna Beloborodova was educated at the Pirogov Russian National Research Medical University, with a degree in pediatrics in 1980, a Ph.D. in 1987, and a specialization in Clinical Microbiology from First Moscow State Medical University in 2004. She has been a Professor since 1996. Currently, she is the Head of the Laboratory of Metabolism, a division of the Federal Research and Clinical Center of Intensive Care Medicine and Rehabilitology, Moscow, Russian Federation. N.V. Beloborodova has many years of clinical experience in the field of intensive care and surgery. She studies infectious complications and sepsis. She initiated a series of interdisciplinary clinical and experimental studies based on the concept of integrating human metabolism and its microbiota. Her scientific achievements are widely known: she is the recipient of the Marie E. Coates Award \\"Best lecturer-scientist\\" Gustafsson Fund, Karolinska Institutes, Stockholm, Sweden, and the International Sepsis Forum Award, Pasteur Institute, Paris, France (2014), etc. Professor N.V. Beloborodova wrote 210 papers, five books, 10 chapters and has edited four books.',institutionString:"Federal Research and Clinical Center of Intensive Care Medicine and Rehabilitology",institution:null},{id:"354260",title:"Ph.D.",name:"Tércio Elyan",middleName:"Azevedo",surname:"Azevedo Martins",slug:"tercio-elyan-azevedo-martins",fullName:"Tércio Elyan Azevedo Martins",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/354260/images/16241_n.jpg",biography:"Graduated in Pharmacy from the Federal University of Ceará with the modality in Industrial Pharmacy, Specialist in Production and Control of Medicines from the University of São Paulo (USP), Master in Pharmaceuticals and Medicines from the University of São Paulo (USP) and Doctor of Science in the program of Pharmaceuticals and Medicines by the University of São Paulo. Professor at Universidade Paulista (UNIP) in the areas of chemistry, cosmetology and trichology. Assistant Coordinator of the Higher Course in Aesthetic and Cosmetic Technology at Universidade Paulista Campus Chácara Santo Antônio. Experience in the Pharmacy area, with emphasis on Pharmacotechnics, Pharmaceutical Technology, Research and Development of Cosmetics, acting mainly on topics such as cosmetology, antioxidant activity, aesthetics, photoprotection, cyclodextrin and thermal analysis.",institutionString:null,institution:{name:"University of Sao Paulo",country:{name:"Brazil"}}},{id:"334285",title:"Ph.D. Student",name:"Sameer",middleName:"Kumar",surname:"Jagirdar",slug:"sameer-jagirdar",fullName:"Sameer Jagirdar",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/334285/images/14691_n.jpg",biography:"I\\'m a graduate student at the center for biosystems science and engineering at the Indian Institute of Science, Bangalore, India. I am interested in studying host-pathogen interactions at the biomaterial interface.",institutionString:null,institution:{name:"Indian Institute of Science Bangalore",country:{name:"India"}}},{id:"329248",title:"Dr.",name:"Md. Faheem",middleName:null,surname:"Haider",slug:"md.-faheem-haider",fullName:"Md. Faheem Haider",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/329248/images/system/329248.jpg",biography:"Dr. Md. Faheem Haider completed his BPharm in 2012 at Integral University, Lucknow, India. In 2014, he completed his MPharm with specialization in Pharmaceutics at Babasaheb Bhimrao Ambedkar University, Lucknow, India. He received his Ph.D. degree from Jamia Hamdard University, New Delhi, India, in 2018. He was selected for the GPAT six times and his best All India Rank was 34. Currently, he is an assistant professor at Integral University. Previously he was an assistant professor at IIMT University, Meerut, India. He has experience teaching DPharm, Pharm.D, BPharm, and MPharm students. He has more than five publications in reputed journals to his credit. Dr. Faheem’s research area is the development and characterization of nanoformulation for the delivery of drugs to various organs.",institutionString:"Integral University",institution:{name:"Integral University",country:{name:"India"}}},{id:"329795",title:"Dr.",name:"Mohd Aftab",middleName:"Aftab",surname:"Siddiqui",slug:"mohd-aftab-siddiqui",fullName:"Mohd Aftab Siddiqui",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/329795/images/system/329795.png",biography:"Dr. Mohd Aftab Siddiqui is an assistant professor in the Faculty of Pharmacy, Integral University, Lucknow, India, where he obtained a Ph.D. in Pharmacology in 2020. He also obtained a BPharm and MPharm from the same university in 2013 and 2015, respectively. His area of research is the pharmacological screening of herbal drugs/natural products in liver cancer and cardiac diseases. He is a member of many professional bodies and has guided many MPharm and PharmD research projects. Dr. Siddiqui has many national and international publications and one German patent to his credit.",institutionString:"Integral University",institution:null}]}},subseries:{item:{id:"7",type:"subseries",title:"Bioinformatics and Medical Informatics",keywords:"Biomedical Data, Drug Discovery, Clinical Diagnostics, Decoding Human Genome, AI in Personalized Medicine, Disease-prevention Strategies, Big Data Analysis in Medicine",scope:"Bioinformatics aims to help understand the functioning of the mechanisms of living organisms through the construction and use of quantitative tools. 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The considerable development of technology, including the computing power of computers, is also conducive to the development of bioinformatics, including personalized medicine. In an era of rapidly growing data volumes and ever lower costs of generating, storing and computing data, personalized medicine holds great promises. Modern computational methods used as bioinformatics tools can integrate multi-scale, multi-modal and longitudinal patient data to create even more effective and safer therapy and disease prevention methods. Main aspects of the topic are: Applying bioinformatics in drug discovery and development; Bioinformatics in clinical diagnostics (genetic variants that act as markers for a condition or a disease); Blockchain and Artificial Intelligence/Machine Learning in personalized medicine; Customize disease-prevention strategies in personalized medicine; Big data analysis in personalized medicine; Translating stratification algorithms into clinical practice of personalized medicine.",coverUrl:"https://cdn.intechopen.com/series_topics/covers/7.jpg",hasOnlineFirst:!0,hasPublishedBooks:!0,annualVolume:11403,editor:{id:"351533",title:"Dr.",name:"Slawomir",middleName:null,surname:"Wilczynski",slug:"slawomir-wilczynski",fullName:"Slawomir Wilczynski",profilePictureURL:"https://s3.us-east-1.amazonaws.com/intech-files/0033Y000035U1loQAC/Profile_Picture_1630074514792",biography:"Professor Sławomir Wilczyński, Head of the Chair of Department of Basic Biomedical Sciences, Faculty of Pharmaceutical Sciences, Medical University of Silesia in Katowice, Poland. His research interests are focused on modern imaging methods used in medicine and pharmacy, including in particular hyperspectral imaging, dynamic thermovision analysis, high-resolution ultrasound, as well as other techniques such as EPR, NMR and hemispheric directional reflectance. Author of over 100 scientific works, patents and industrial designs. Expert of the Polish National Center for Research and Development, Member of the Investment Committee in the Bridge Alfa NCBiR program, expert of the Polish Ministry of Funds and Regional Policy, Polish Medical Research Agency. 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