Studies reporting the use of natural molecules and Pickering stabilizers selected from the retrieved 788 documents of the bibliometric search.
\r\n\tThis book intends to provide the reader with a comprehensive overview of the current state-of-the-art novel imaging techniques by focusing on the most important evidence-based developments in this area.
",isbn:null,printIsbn:null,pdfIsbn:null,doi:null,price:0,priceEur:0,priceUsd:0,slug:null,numberOfPages:0,isOpenForSubmission:!0,isSalesforceBook:!1,isNomenclature:!1,hash:"d9159ce31733bf78cc2a79b18c225994",bookSignature:"Dr. Gabriel Cismaru",publishedDate:null,coverURL:"https://cdn.intechopen.com/books/images_new/11867.jpg",keywords:"Hypertrophic Cardiomyopathy, Dilated Cardiomyopathy, Restrictive Cardiomyopathy, Transesophageal Echocardiography, Intracardiac Echocardiography, 3-Dimensional Echocardiography, Adult Congenital Heart Disease, Tetralogy of Fallot, Transposition of the Great Vessels, Coronary Artery Disease, Risk Stratification, Revascularization",numberOfDownloads:null,numberOfWosCitations:0,numberOfCrossrefCitations:null,numberOfDimensionsCitations:null,numberOfTotalCitations:null,isAvailableForWebshopOrdering:!0,dateEndFirstStepPublish:"April 21st 2022",dateEndSecondStepPublish:"May 19th 2022",dateEndThirdStepPublish:"July 18th 2022",dateEndFourthStepPublish:"October 6th 2022",dateEndFifthStepPublish:"December 5th 2022",dateConfirmationOfParticipation:null,remainingDaysToSecondStep:"3 months",secondStepPassed:!0,areRegistrationsClosed:!0,currentStepOfPublishingProcess:4,editedByType:null,kuFlag:!1,biosketch:"Dr. Cismaru Gabriel is an Assistant Professor at the University of Medicine and Pharmacy Cluj-Napoca, certified in Cardiology. After completing his certification in cardiology, Dr. Cismaru began his electrophysiology fellowship at the Institut Lorrain du Coeur et des Vaisseaux Louis Mathieu. He has authored or co-authored peer-reviewed articles and book chapters in the field of cardiac pacing, defibrillation, electrophysiological study, and catheter ablation.",coeditorOneBiosketch:"Raluca Tomoaia is an MD, Ph.D. in novel techniques in Echocardiography at the University of Medicine and Pharmacy in Cluj-Napoca, Romania., assistant professor, and a researcher in echocardiography and cardiovascular imaging.",coeditorTwoBiosketch:null,coeditorThreeBiosketch:null,coeditorFourBiosketch:null,coeditorFiveBiosketch:null,editors:[{id:"191888",title:"Dr.",name:"Gabriel",middleName:null,surname:"Cismaru",slug:"gabriel-cismaru",fullName:"Gabriel Cismaru",profilePictureURL:"https://mts.intechopen.com/storage/users/191888/images/system/191888.png",biography:"Dr. Cismaru Gabriel is an assistant professor at the Cluj-Napoca University of Medicine and Pharmacy, Romania, where he has been qualified in cardiology since 2011. He obtained his Ph.D. in medicine with a research thesis on electrophysiology and pro-arrhythmic drugs in 2016. Dr. Cismaru began his electrophysiology fellowship at the Institut Lorrain du Coeur et des Vaisseaux Louis Mathieu, France, after finishing his cardiology certification with stages in Clermont-Ferrand and Dinan, France. He began working at the Rehabilitation Hospital\\'s Electrophysiology Laboratory in Cluj-Napoca in 2011. He is an experienced operator who can implant pacemakers, CRTs, and ICDs, as well as perform catheter ablation of supraventricular and ventricular arrhythmias such as ventricular tachycardia and ventricular fibrillation. He has been qualified in pediatric cardiology since 2022, and he regularly performs device implantation and catheter ablation in children. Dr. Cismaru has authored or co-authored peer-reviewed publications and book chapters on cardiac pacing, defibrillation, electrophysiological studies, and catheter ablation.",institutionString:"Iuliu Hațieganu University of Medicine and Pharmacy",position:null,outsideEditionCount:0,totalCites:0,totalAuthoredChapters:"7",totalChapterViews:"0",totalEditedBooks:"2",institution:{name:"Iuliu Hațieganu University of Medicine and Pharmacy",institutionURL:null,country:{name:"Romania"}}}],coeditorOne:null,coeditorTwo:null,coeditorThree:null,coeditorFour:null,coeditorFive:null,topics:[{id:"16",title:"Medicine",slug:"medicine"}],chapters:null,productType:{id:"1",title:"Edited Volume",chapterContentType:"chapter",authoredCaption:"Edited by"},personalPublishingAssistant:null},relatedBooks:[{type:"book",id:"5970",title:"Bedside Procedures",subtitle:null,isOpenForSubmission:!1,hash:"ba56d3036ac823a7155f40e4a02c030d",slug:"bedside-procedures",bookSignature:"Gabriel Cismaru",coverURL:"https://cdn.intechopen.com/books/images_new/5970.jpg",editedByType:"Edited by",editors:[{id:"191888",title:"Dr.",name:"Gabriel",surname:"Cismaru",slug:"gabriel-cismaru",fullName:"Gabriel Cismaru"}],productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"9064",title:"Epidemiology and Treatment of Atrial Fibrillation",subtitle:null,isOpenForSubmission:!1,hash:"1cd6bf2b3181eb82446347fbe478a2bc",slug:"epidemiology-and-treatment-of-atrial-fibrillation",bookSignature:"Gabriel Cismaru and Keith Andrew Chan",coverURL:"https://cdn.intechopen.com/books/images_new/9064.jpg",editedByType:"Edited by",editors:[{id:"191888",title:"Dr.",name:"Gabriel",surname:"Cismaru",slug:"gabriel-cismaru",fullName:"Gabriel Cismaru"}],productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"6550",title:"Cohort Studies in Health Sciences",subtitle:null,isOpenForSubmission:!1,hash:"01df5aba4fff1a84b37a2fdafa809660",slug:"cohort-studies-in-health-sciences",bookSignature:"R. 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In 1876, Henry John Horstman Fenton first discovered the enhanced oxidizing power of ferrous ions (Fe+2), hydrogen peroxide (H2O2) on tartaric acid (C4H6O6). When Fenton added sodium hydroxide (NaOH) to the mixture, the solution became bright purple [1]. Fenton made a decision to find out what that purple molecule was. That goal became his career and immortalized his name in the annals of chemistry [2].
Eighteen years later, Fenton repeated the experiment, again adding to a tartaric acid solution, a catalytic amount of FeSO4, followed by H2O2 with the molar ratio of each factor: C4H6O6 / H2O2 / Fe+2 = 1.0: 1.0: ‘catalytic’. Fenton then isolated the reaction product by sequentially precipitating the acid with heavy metal ions, weighing the salt to calculate the molar formula, re-purifying the acid, then repeating the process with a different cation, thus calculating the empirical formula of the new acid. The new acid bound one divalent cation ion or two monovalent cations ions per molecule, and thus was a di-acid. Fenton determined: 1) the molecule had the empirical formula C4H4O6; 2) was a 4-carbon di-acid; 3) produced by abstraction of two hydrogen atoms from tartaric acid [3].
Fenton (1896) assumed that the 4-chain backbone was not severed, limiting his options to three possible structures: 1) loss of two hydrogen from a single middle carbon, forming a hydroxy-, keto-, di- carboxylic acid: (2-hydroxy-3-oxosuccinic acid); or 2) loss of two hydrogen from the internal adjacent carbon atoms forming a double bond with the HO- groups either in: 2a)
The first structure was eliminated when the di-acid failed to form a hydrazone with phenylhydrazine (an aldehyde/ketone reactive agent). The assumption that the molecule had two internal hydroxyl groups was verified when the molecule formed a 4-carbon di-ester, di-anhydride with either acetyl chloride or acetic anhydride.
The structure of the molecule was finalized by reaction with aniline. Fenton knew (from literature) that the 1:1 product of aniline and fumaric acid (C4H2O4: -C2H = HC3- in
Following the initial discovery, Fenton tested the range of his new reagent. Fenton & Jackson (1899) oxidized aliphatic alcohols, polyalcohols, and benzoic, then FeSO4 followed by H2O2, added stepwise, in molar ratios of (1: 0.1–0.25: 1), adding the peroxide gradually in small amounts.
The aliphatic alcohols: (CH3OH, CH2CH5OH,
On the other hand, the polyalcohols (C2H6O2, C3H8O3, C4H10O4, C5H12O5, and C6H14O6) showed temperature increases and release of gases, whereas the H2O2 only controls showed no reaction for either group. When the oxidized polyalcohols were reacted with phenylhydrazine forming osazones, indicating that hydroxyl groups of the polyols were oxidized to carbonyls, forming aldoses and/or ketoses. The oxidation of hydroxyl groups in the polyalcohols to aldehydes or ketones required a loss of two hydrogen atoms (H•) from the molecular formula.
Benzoic acid (C7H6O2) was oxidized to salicylic acid (
Fenton & Jones (1900) repeated testing the oxidizing abilities of Fe+2/H2O2 on a larger set of aliphatic and polyhydroxy acids. Their method was to prepare a 1 M solution of reagent in H2O at 0°C, add FeSO4 to 0.125 M, then add H2O2 to 0.25 M (-FeSO4 was the control). The authors again reported that aliphatic acids appeared non-reactive while polyhydroxy acids showed vigorous and energetic reactions. The oxidized acids reacted with phenylhydrazine, and the precipitates were purified by crystallization, confirming that hydroxyl groups were oxidized to ketones or aldehydes and the molecules identified by melting point determinations. The oxidation of benzoic acid to salicylic acid was also confirmed [7].
Collectively, Fenton (1896) and Fenton et al. (1899, 1900) presented evidence for three different reactions: 1) carbon–carbon double bond formation with loss of 2 H-; 2) carbon–oxygen double bond formation with loss of 2 H- (both aldehydes and ketones; and 3) oxygen addition to phenol.
Following Fenton’s lead, Cross, Bevan, & Smith (1898) oxidized glucose with FeSO4 / H2O2 and isolated “glucosone” (2-keto-, glucose). The experiment consisted of: 1) in 100 mL H2O, 4% or 10% glucose; 2) FeSO4 added to a concentration of 1/104 ratio with glucose, and 3) H2O2 was gradually added to a final 1:1 molar glucose/ H2O2 with stirring on ice [8]. Theorizing that 2-keto-, glucose would be indigestible, the authors used yeast to scavenge unoxidized glucose. After filtering out the yeast, the authors found that the solution still contained carbonyl molecules, as indicated by reduction of CuO. The solution retained 20% of the reducing power of the original glucose solution. The oxidized glucose solution also increased in acidity. After drying (105°F / 40.6°C), the dried residue comprised 88% of the weight of glucose including 3.8% furfural.
The solids were resuspended in chilled water and reacted with phenylhydrazine (PHZ), a reversible carbonyl-reactive label). The rationale was that while glucosone and glucose are likely to be equally soluble in most solvents, the double substituted glucosazone was expected to have a different solubility profile from the gluco-hydrazone. The glucosone reacted with 2 moles of PHZ, whereas glucose reacts with only one mole of PHZ, therefore their solubility in differences in organic solvents would be greater than the unlabeled molecules. After purification, PHZ-labeling was reversed with H2SO4, allowing analysis of purified glucosone. The authors repeated their method with fructose and sucrose recovering only glucosone from all three sugars implying: 1) fructose oxidized to glucosone; and 2) sucrose hydrolyzed to glucose and fructose.
Morell
Morrell et al. (1899b) increased glucose to glucosone yields by: 1) slow addition of H2O2, 2) controlling temperature with refrigeration, 3) controlling pH and precipitating organic acid with (Pb(OAc)2). This method increased glucosone yield to 10%. Morell et al. purified PHZ-glucosazone and PHZ-mannosazone from their corresponding PHZ-hydrazones, but were not able to purify PHZ-arabinosazone, PHZ-rhamnosazone, and PHZ-galactosazone from their PHZ-hydrazone contaminants [10].
Morrell
Morrell
Morell et al. (1903) oxidized aldoses with Fenton’s reagent, precipitate organic acids with Pb(OAc)2 and Ba(OH)2, then label the oxidation mixture for carbonyl groups with methyl-, phenyl-hydrazine (MPHZ). However, galactose and arabinose MPHZ-osazones were not separable from their MPHZ-hydrazones. The authors tested bromo-, phenylhydrazine (BPHZ) and found that BPHZ-arabinosazone was easily separable from BPHZ-arabinose hydrazone using benzene as the solvent [13].
Morrell & Bellars (1905) retested purification of all the aldose osazones with BPHZ. BPHZ-labeling after Fenton oxidation allowed sharp separations of BPHZ -osazones from the corresponding hydrazones in benzene, thus achieving the goal of acquiring pure osazones after Fe+2/H2O2 oxidation [14]. [Considering that both Cross & Bevan (1898) and Morell et al. (1899) stated that the purified osones were tasted, the implication is that both groups were investigating the osones as non-caloric sweeteners].
Cross & Bevan (1898) surveyed the by-products yielded by Fenton oxidation of aldoses to glucosones. The secondary products included: tartronic acid, (∼ 8%), acetic acid (∼5%), formic acid (∼15%), and furfural(s) (∼ 4%). Missing carbohydrate mass was assumed to be lost as carbon dioxide (CO2). The authors noted that Fenton oxidation of glucose produced furfural, but fructose did not; on the other hand, glucose produced lower amounts of dicarboxylic acids and pentoses [8]. Cross & Bevan (1899) oxidized a 2% solution of furfural with H2O2 and a catalytic amount of FeSO4. Formic, acetic acid, and a red precipitate identified as pyromucic acid were isolated. The authors also reported that Fe+2/H2O2 oxidized benzene to phenol, followed by additional hydroxylations [15].
Morrell et al. (1903) used Pb(OAc)2 and Ba(OH)2 to precipitate the sugar acid impurities in the quest to purify the osones. From the oxidation of glucose and fructose, the experimenters recovered the several polyhydroxy acids after precipitation with Pb+2 or Ba+2 ions. After removal of Pb+2 and Ba+2 ions with H2SO4, the solubilized acids were identified as glycolic and oxalic. The Pb+2 / Ba+2 soluble acids were separately precipitated as calcium salts and identified as glyoxylic and trihydroxy-butyric acids [13].
In sum, Cross & Bevan (1898, 1899) and Morrell et al. (1899–1903) confirmed that Fenton’s reagent was responsible for the following reactions: dehydrations producing C=C bonds, C-C bond cleavages, and oxygen atom additions forming hydroxyl, aldehyde, ketone, and carboxylic acid groups, creating new classes of organic molecules.
Prof. Henry J. H. Fenton died in 1929 without knowing the mechanism of the reagent that he discovered. Within three years of his death, two competing mechanisms naming two different intermediate molecules were published.
In 1932, Fritz Haber & Joseph Weiss (1932) published in
Hydroxyl radical reactions.
The authors proposed that the Fe+2 ion donates an electron to the peroxide molecule, cleaving the O-O bridge producing a hydroxyl radical (HO•) and a hydroxide ion (HO−). The hydroxyl radical then attacks another peroxide molecule, forming superoxide, eventually generating oxygen [16, 17].
On the other side of the Atlantic Ocean, William Bray & H. M. Gorin published in Journal of the American Chemical Society (1932) that oxygen production after addition of Fe+2 ions to H2O2 in H2O is due to creation of the ferryl-oxo ion (Fe = O)+2 (Figure 2). After creation, the ferryl-oxo ion then reacts with another ferryl-oxo molecule to produce oxygen gas, recycling the ferrous ion. The authors proposed that an oxygen atom (•O•) is abstracted by a Fe+2 ion from the peroxide molecule, forming the ferryl-oxo ion (Fe = O+2) (no net change in charge) and H2O [18].
Ferryl-Oxo ion reactions.
[Bray & Gorin named the molecule ‘ferryl ion’, but the term is currently used for the Fe+4 ion (without oxygen) [19]. To avoid confusion, the Bray & Gorin molecule is named here ‘ferryl-oxo’ ion. For similar reason, Barton’s ‘perferryl ion’ will be named ‘perferryl-oxo ion’].
These two papers divided the scientific community into partisan camps with sports-like fanaticism that continues today. Champions of the hydroxyl radical theory include: JD Rush, WH Koppenol [20, 21, 22, 23], C. Walling [24, 25, 26], M. Kremer [27, 28, 29], JH Merz, WA Waters, [6, 30, 31], as well as many others. The scientists who argued for the existence of the ferryl-oxo ion included JT Groves [32, 33, 34, 35], DA Wink [36, 37, 38], and DT Sawyer [39], among many others.
Only exceeded by a fluorine (F0) atom, the powerful hydroxyl (HO•) radical is the second strongest electrophile, and will even oxidize chlorine ion(s) (Cl−) to elemental chlorine (Cl0) or gas (Cl2) [40]. A hydroxyl radical will strip an electron from an element (except F
Hydroxyl radicals (HO•: Figure 1) are created by:
Donation of an electron to H2O2 from a transition metal ion (Figure 1a) (with
Splitting H2O2 with UV (λ = 253.7 nm) or γ- radiation (from 60Co) produces two hydroxyl radicals:
(See (Eq. (2)) (Figure 1d) [42];
UV flash irradiation of oxygen donating molecules, such as N2O to create oxygen atoms (•O•) that react with water molecules:
Ionizing water with electrically produced β-rays to produce radicals
(H• radicals likely combine with each other, as also H− & H+ ions; thus escaping as H2 gas) [42, 45].
Once created, hydroxyl radicals (HO•) will oxidize an element, ion, or compound, by extracting an electron to form HO− and a cation, increase the valence of another ion by +1, or abstract H- from an X-H bond of organic molecule, forming H2O and an organic radical (Figure 1e).
A hydroxyl radical abstracts H• from an alkane to create a C• radical and water (Figure 1e) [46, 47]. A second HO• is required to collide with C• to form an alcohol (Figure 1f), thus a high HO•/ substrate ratio is required to produce alcohols. Subsequent HO• hydroxyl attacks to the same carbon atom progressively oxidizes and adds oxygen atoms, producing aldehydes/ketones, then organic acids, and finally, carbon dioxide [31, 42, 48]. An example of a hydroxyl radical reaction sequence for oxidation of methane:
There is no guarantee that a second HO• will collide with a carbon radical to make an alcohol.
Under low HO• concentrations, long-lived hydrocarbon radicals fuse to each other to make large complex hydrocarbons via R1C••CR2 fusions (Figure 1g). The hydroxyl radical oxidation of methane can also follow:
Thus carbon–carbon fusions are a hallmark of hydroxyl radical reactions [42, 49].
The HO• radical is: 1) uncharged, and 2) will abstract an electron any atom (except F−) or H• from a molecule it collides with, thus the (HO•) radical is an indiscriminant oxidant. Its oxidation profile determined by accessibility and rate of diffusion. In the oxidation of simple alkanes, the oxidation preference is: 1° H > 2° H > 3 H°.
Hydroxyl radicals (HO•) [17] can be created by H2O2 receiving an electron from a transition metal ion [41], or by splitting an oxygen donating molecule, either H2O2, with UV light or radiation [42, 50] or N2O (aq.) with UV light [43, 44]. Hydroxyl radicals can be quenched/scavenged by reducing agents [51] including aliphatic alcohols [50], DMSO [52], acetate ions [22], polyols [53], H2S [54], and NO [55]. These reagents are included as radical traps where HO• radical oxidations are undesirable.
Waters (1946) reported that HO• radical oxidation of ethylene glycol (CH2(OH)-CH2(OH)) produced both glycoaldehyde (CH(O)-CH2(OH)) and formaldehyde (2x: CH2O). To determine which H• abstraction caused C-C bond cleavage, Waters oxidized pinacol, which has no available O-
Droege & Tully (1986a,b) oxidized gaseous ethane (1H, 2H, and mixed) (46)] and propane (1H, 2H, and mixed) [47] with UV-activated N2O & H2O to compare oxidation rates of the terminal vs. center carbons, and test the isotope effect on HO• oxidation for different positions of the ethane and propane molecules. The authors found that there was no difference in positional abstraction for hydrogen vs. deuterium at 1° (ethane & propane) or 2° positions (propane only); however C-C chain fusions increased with temperature.
Baxley & Wells (1998) oxidizing tertiary alcohols with HO• radicals in air. HO• radicals were generated by UV activation of CH3ONO, NO, and O2 gases. H-abstraction from the sole -OH group caused C-C cleavage producing a ketone, a hydrocarbon and water. Abstractions from C-H bonds produced either addition of a second hydroxyl group or fusions producing long chain diols, however the authors noted that the hydroxyl group of 2-butanol was targeted more frequently than of 2-pentanone [48].
Gilbert and King (1981, 1984) oxidized glucose with HO• generated by Ti+3/H2O2. Using electron spin resonance (ESR) the authors concluded that HO• radical produced carbon (C•) radicals at all positions in equal ratios, indicating distributed attack by HO• toward all carbon positions in glucose, the established signature of HO• oxidations [57, 58].
Dizdaroglu & Von Sonntag reacted glucose [43] and cellobiose (44) with HO• generated from UV irradiation of N2O saturated H2O. By mass spectroscopy, the authors identified several 6-carbon derivatives of glucose including gluconic and glucuronic acids, several hexosuloses, hexodialdose, and. Several dehydro-hexoaldoses, proposing that addition of H• or HO• radicals occurred after abstraction of –H or –OH groups from glucose. The authors determined for both carbohydrates, all carbons were oxidized equally.
In addition to 6-carbon molecules, Von Sonntag and coworkers reported fragmentation of glucose into various aldoses, formaldehyde, formic acid, carbon dioxide, and carbon monoxide, representing different C-C bond cleavages. The authors did not explain the origins of the C-C bond cleavage products.
In summary, the hydroxyl radical (HO•) is a powerful but non-selective oxidant. It can abstract electrons from any molecule or element with exception of fluorine. Hydroxyl radicals will abstract hydrogen atoms (H•) from organic molecules from any accessible C-H, O-H, or N-H (59) bond at rates proportional to accessibility by simple diffusion [43, 46, 57].
The ferryl-oxo [(Fe = O)+2] ion, a less powerful oxidant than the HO• radical, is created by oxygen abstraction from H2O2 (Figure 2a). The oxidizing power of the (Fe = O)+2 ion is moderately stronger than the strength of C-H bond of an alkane and roughly equivalent to the C-H bond strength of benzene; the (Fe = O)+2 ion is reported not to abstract H• from anhydrous acetonitrile (CH3-C ≡ N) [59]. Though weaker than the HO• radical, the (Fe = O)+2 ion is a discriminatory oxidant, abstracting H from the weakest X-H bond in a molecule and oxidizing molecules with the weakest X-H bonds in a mix of molecules (Figure 2b) [32, 33, 34, 35]. In the oxidation of simple alkanes, the oxidation preference is: 3° H > 2° H > 1° H (Figure 2).
Groves & coworkers demonstrated that oxidation of alkanes by (Fe = O)+2 in non-aqueous environments produced alcohols without carbon–carbon fusions. Addition of -OH groups to alkanes was both
Groves et al. proposed a two-step mechanism to explain their results (Figure 2b):
The ferrous ion abstracts oxygen from peroxide forming the ferryl-oxo ion with a coordinate double bond;
The ferryl-oxo ion [(Fe = O)+2] abstracts H• from a C-H bond creating a C• radical and ferric hydroxide [(Fe+3OH) or (Fe-OH)+3] (Figure 2b);
The oxygen of ferric hydroxide (Fe+3OH) reacts with the C• radical, producing an alcohol (RC-OH), and regenerating the Fe+2 ion.
Unlike HO•, the [Fe = O]+2 is both stereo- and regio- selective. For hydrocarbons, the H-C oxidation preference order is: 3° C-H > 2° C-H > 1° C-H. The basis of the rebound effect is likely due to attraction of the electrophile C• radical and the nucleophile oxygen (•O•) of the (Fe-OH)+3 ion. The oxygen is added to the same bond position on the oxidized carbon.
[Fenton’s original reaction: (Eq. (7)) violates Groves’ model because tartaric acid oxidation follows a different pathway:
Erik Hückel’s double bond resonance theory states that molecules with 4N + 2 unpaired electrons in conjugated (staggered) double bonds are extraordinarily stable. In the oxidation of tartaric acid, the abstraction of the first H• from C2 is followed by ejection of the second H• from C3 to form a C=C bond, because the central C=C bond is conjugated to the flanking C=O bonds of the terminal carboxylic acids. Thus, Fenton’s molecule was resistant to further oxidations, allowing him to discover it].
Ferryl-oxo ion [(Fe = O)+2] oxidation of oxygen-containing organic molecules behaves differently from hydrocarbon oxidations (Figure 2c and d). Carbon and hydrogen have similar affinity for electrons, therefore the electron pair is shared equally and in a hydrocarbon, hydrogen-carbon all bonds are about equal strengths. Oxygen (O) heteroatoms have a higher affinity for electrons than carbon or hydrogen atoms, making the C-O bond stronger than a C-C bond, while weakening other bonds extending from the hydroxyl carbon significantly [19, 60, 61, 62].
As an example, when (HO•) oxidizes ethanol, H• abstraction occurs indifferently from any of the six C-H positions, producing roughly 50% ethylene glycol and 50% acetaldehyde yield. On the other hand, when ethanol is oxidized by (Fe = O)+2 ion, the bond strengths of the methyl C-H bonds are ∼96 kCal/mole, whereas the hydroxyl C-H bond strengths are ∼81.6 kCal/mol and the O-H bond strength is ∼104 kCal/mole (60). Because the (Fe = O)+2 ion has the higher probability of abstracting H• from a hydroxyl carbon (due to bond strength) or hydroxyl oxygen (due to charge attraction) rather methyl carbons, acetaldehyde will be formed in preference to ethylene glycol [60, 63, 64, 65, 66].
Following the Coon & White (1977) discovery of the Fe+3-heme core in cytochrome P450 and its ability to sever the O-O bond, and oxidize NADPH2 [67, 68], Okamoto et al. (1985) mimicked the ability of enzyme P450scc to split a C-C bond of a diol in 1-, 2-, bis-(4-methoxyphenyl)ethane-l,2-diol using Fe+3 ion, O2, and a reductant (N-benzyl-3-carbamoyl-1,4-dihydropyridine) [69].
Okamoto et al. (1988) found that Fe+2 + H2O2 could substitute for Fe+3 and O2 to cleave diols to paired aldehydes. Using various inhibitors and/or substituting ferric for ferrous ion, the authors concluded that (Fe = O)+2 was created and was the oxidant that cleaved the 1-, 2-, diols (Figure 2c). The authors also discovered that when one hydroxyl group was substituted, paired aldehydes formed, but when both hydroxyl groups were blocked, no aldehydes were produced (Figure 2d) [70].
Sugimoto and Sawyer (1985a) reported that Fe+2 and two moles of H2O2 oxidized alkenes (hydrocarbons with double bonds) to paired aldehydes formed by (Fe = O)+2. The authors proposed that Fe+2 ion and H2O2 caused dioxygen addition to a double bond, forming a dioxetane, that then scissioned to a diol; a second oxidation (Fe = O)+2 scissioned the diol to paired aldehydes. The authors saw similar oxidative behavior when CH3-O-O-H and
Thus 1-, 2-, diols produce the same products when oxidized by either HO• (56) or (Fe = O)+2 [68] oxidants indicating that the formation of paired aldehydes is faster than oxygen addition reactions of alkanes. [Though contemporary, the Sawyer’s and Oka’s research teams did not appear to be aware of each other, or of Waters (1946)].
The rationale for asymmetric cleavage of diols (Figure 2c and d) is due to the additive weakening of the C-C bond between the two hydroxyl groups [19, 60, 61, 65]. When H• is abstracted from a hydroxy oxygen of a diol pair, the weakest bond of the oxygen-centered (H-C-O•) carbonyl radical is the C-C bond between the diol pair (•O-R1CH ∼ R2CH-OH); electron abstraction from the C ∼ C bond produces paired aldehydes (Figure 2c). However, when H• is abstracted from a C-H bond of a hydroxyl carbon, the weakest bond of the diol group is the O-H bond opposite the C• radical (H ∼ O-R1C•); the hydrogen atom is ejected from carbon-centered (H ∼ O-C•) carbonyl radical to form the carbonyl bond. Abstraction of H• from a tertiary -OH group can cause ejection of a C• radical to form the C=O bond (Figure 2d) [60, 61, 62, 63, 64, 65, 66].
[Fenton’s oxidation of tartaric acid should have produced two products: 2-, 3-, dihydroxy-, maleic acid [HOOC-C(OH) = C(OH)-COOH]
Though HO• radical and (Fe = O)+2 ion both create a C• radical, the differences between the two oxidants is: 1) HO• is a 1 e− oxidant, whereas (Fe = O)+2 ion is a 2 e− oxidant, thus two independent HO• oxidations are required to make a hydroxyl group; and 2) reducing agents that trap HO• radicals and thus halt HO•-based oxidations, do not disrupt ferryl-oxo ion oxidations. The most likely explanation radical quenching by the ferryl-oxo ion is the proximate distance of Fe+3-O-H and C• radical is coupled with likely nucleophile / electrophile attraction, allowing rapid re-reaction to occur [36, 37, 38].
The noted crypto-HO• positional effect seen in Fe+2/H2O2 catalyzed oxidations is likely due to localized binding of Fe+2 ions to a substrate that has O heteroatoms when it added to the substrate prior to H2O2 [22, 23, 72] addition.
Fenton conducted Fe+3/H2O2 experiments but did not note any reactions and assumed that no reaction(s) had occurred [5, 7]. However, on the other side of the English Channel, Fenton’s contemporaries found contrary evidence.
Spring (1895) mixed H2O2 solutions with different pure chemical substances noting which substances caused oxygen gas release. Spring noted that both ferrous and ferric chlorides decomposed H2O2 and released oxygen gas [40].
Ruff (1898) used basic ferric acetate and H2O2 to oxidize gluconic acid to arabinose and carbon dioxide, a C1-C2 bond cleavage with oxidations of both C1 and C2, the reaction now known as ‘Ruff’s degradation’ [73].
Bohnson (1921) noted that a solution of a ferric salt in water, dilute enough to show only very slight color, turns brightly lavender with ‘1 or 2 drops’ of 30% H2O2 followed by O2 gas release from the solution. After bubbling ends, no residual H2O2 remained in the solution, indicating complete decomposition. The author observed that when H2O2 is added to Fe+3 salts, a lavender color appears briefly. Bohnson speculated that the color represented a transient higher Fe oxidation state. Bohnson trapped the lavender pigment with cold KOH coloring the solution red, then Ba(OH)2, forming a red gelatinous precipitate. Washing the precipitate with HCl released chlorine gas. Bohnson determined the empirical formula of the precipitate: barium ferrate (BaFeO4), thus isolating the Fe+6 oxidation state as FeO4−2 (ferrate) ion. Bohnson also prepared potassium ferrate (K2FeO4) by bubbling chlorine gas through a solution of Fe(OH)3/KOH solution, producing a deep lavender color; addition of Ba(Cl)2 to the lavender solution again formed the red precipitate: BaFeO4 [74].
Bohnson (1921) also demonstrated direct conversion of ethanol to acetic acid. Bohnson noted that addition of Fe+3 ions to an H2O2 solution produced oxygen gas, but addition of ethanol to the H2O2 solution prior addition of Fe+3 ions disrupted oxygen evolution, leading the author speculated that ethyl alcohol was oxidized to acetaldehyde or acetic acid. Bohnson also compared of oxidation by ethanol by Fe+2/H2O2 vs. Fe+3/H2O2 and found that Fe+2/H2O2 produces acetaldehyde, then acetic acid, whereas Fe+3/H2O2 oxidized ethanol to acetic acid primarily, with only trace amounts of acetaldehyde detected. Bohnson proposed that Fe+3/H2O2 oxidized ethanol directly to acetic acid, bypassing acetaldehyde formation [74].
Walton & Christensen (1926) compared the oxidation of ethanol with FeSO4/H2O2 or Fe2(SO4)3/H2O2 under anhydrous conditions. Separately assaying for acetaldehyde and acetic acid, the authors noted that when ethanol is oxidized with FeSO4/H2O2 acetaldehyde appeared before acetic acid, whereas when ethanol is oxidized by Fe2(SO4)3/H2O2 acetic acid appears long before acetaldehyde, proving that Fe+3/H2O2 oxidation exhibits non-Fenton-like behavior, thus confirming Bohnson (1921) [75].
Wieland & Franke (1928) reported that under strong acidic conditions Fe+3/H2O2 oxidized formic acid (HCOOH) to CO2 and H2O, and dihydroxymaleic acid (HOOC-(OH)C=C(OH)-COOH) to 2,3 dioxo-propanoic acid (HOOC-C(O)-C(O)-COOH) and CO2 [76].
Goldschmidt & Pauncz (1933) investigated the Fe2(SO4)3/H2O2/ethanol reaction and confirmed that ethanol was oxidized directly to acetic acid. The authors also explained that Fenton & Jackson (1899) and Fenton & Jones (1900) did not detect aldehydes from aliphatic alcohols because the 1:1 molar ratio of H2O2 and alcohol was sufficient to oxidize all the alcohol to organic acids [77].
Even as late as 1989, Fe+3/H2O2 oxidation articles appeared noting unusual oxidations. Sanderson et al. (1989) submitted a patent for co-synthesis of
White & Coon (1977) summarized the discovery of the mechanism of respiration by mitochondrial enzyme cytochrome P450. Cytochrome P450 uses a Fe+3 ion chelated in a heme ring to conduct the reduction: (Eq. (8)) [67, 68].
Responding to the discovery that the critical enzyme of respiration forms a Fe+3 = O intermediate to split the dioxygen molecule, Barton et al. (1983) sought to mimic the biological reaction using chelated Fe+3 ions and peroxide ion (O2−2) instead of oxygen as a new process to oxidize hydrocarbons to alcohols. Working with alkanes (R1-CH2-R2), Barton expected that pyridine-chelated Fe+3 ions and potassium peroxide (K2O2) would produce alcohols (Figure 3) [54].
Perferryl-Oxo ion reactions.
What Barton did not expect was that the reaction produced a mix of alcohols [R1-HC(OH)-R2]
To understand the reaction mechanism, Barton and co-workers studied the oxidation of adamantine (C10H16, 4 tertiary, 6 secondary, 0 primary C-H groups). Despite the preponderance of secondary carbons, Barton’s reactant favored oxidation of tertiary vs. secondary carbons by a 5:1 margin indicating that the oxidant behaved similar to the ferryl-oxo ion, but single step ketone addition was never reported for (Fe = O)+2 oxidations [54].
Sugimoto & Sawyer (1985b) confirmed and extended Barton’s findings by using Fe+3 and H2O2 to oxidize hydrocarbons molecules with double and triple bonds, isolating epoxides (R1-C-O-C-R2) and oxetanes (R1-C-O−O-C-R2) [79].
Seven years elapsed until Barton and coworkers resolved the perferryl-oxo structure and oxidation mechanism (Figure 3).
Couching their model on the accepted behavior of Fe+3 nucleus of cytochrome P450 [80, 81, 82], Barton et al. (1990) proposed (Fe = O)+3 as the reaction product of Fe+3 and H2O2 or Fe+2 and O2• − (superoxide anion) (Figure 3a). [Barton et al. (1990–8) wrote the structure of the perferryl-oxo ion as [FeV=O]. The (FeV=O(−2)) and (Fe = O)+3 formulas are equivalent for atoms, bonds, and net charge].
Barton et al. (1990): 1) proposed a bifurcated pathway leading either to alcohol or ketone formation, showing that the alcohol/ketone ratio could be varied with addition of dianisyl telluride, and 2) determined that both alcohol and ketone formation occurred in two steps, choosing different non-reversible paths at the second reaction [83, 84].
Barton and Doller (1992) mapped out steps of the pathway of perferryl-oxo ion oxidation of hydrocarbons (Figure 3b–d):
Step 1 (Figure 3b): Formation of Fe+4-C-R intermediate. Using diphenyl-diselenide (Ph-Se-Se-Ph), or phenyl selenol (Ph-Se-H), Barton trapped the Fe+4-C-R intermediate as a stable (Fe+3-Ph-Se-C-R) intermediate as detected by mass spectroscopy (structure not specified).
Step 2 (Figure 3c): Oxygen Insertion to form Fe+3-O-O-C-R intermediate. Comparing 16O2 and 18H2O2, the authors detected primarily 16O-labeled alcohols and ketones indicating that O2 (not O2−2) formed the dioxygen bridge. The authors proposed that in an anoxic environment, peroxide is oxidized to dioxygen by ferric ions in sufficient quantities to complete the reaction as follows: [Eq. (9)]
Step 3 (Figure 3d (1 & 2)): Bifurcated Pathways Arise from Differential Cleavage of the O-O Bridge. The Fe+3-O-O-C-R intermediate is the branch point between the 2e- and 4e- oxidative pathways: a) scission of the Fe+3-O-|-O-R bond produces an alkoxide (R-O―) and the (Fe = O)+3 ion (Figure 3d.1); b) scission of the Fe+3-|-O-O-R bond produces Fe+3 ion and a peroxyl (―O-O-R) ion which then degrades to a ketone (R = O), and an oxide ion (O−2) (Figure 3d.2).
Barton and Doller (1992) trapped the ferric-peroxy-carbon (Fe+3-O-O-C-R) cleavage intermediates with tri-methoxy phosphine (P(OMe)3). P(OMe)3 reacted with either oxygen (R-C-O*-O-Fe+3 or R-C-O-O*-Fe+3) trapping potential oxygen bridge cleavage products as R-C-O-P(OMe)3 and R-C-O-O-P(OMe)3 respectively. Thus Barton and Doller (1992) explained the mechanism of bifurcated production of alcohols or ketones from alkanes by perferryl-oxo (Fe = O)+3 ion (85). Barton’s oxidation scheme was confirmed by Schuchardt et al. (2001) [55].
Barton’s perferryl-oxo ion oxidation theory explains Ruff’s oxidation gluconic acid to arabinose (1898) [71] the one-step conversion of ethanol to acetic acid observed by Bohnson (1921) [72], Walton & Christensen (1926) [75], Goldschmidt & Pauncz (1933) [75], and the co-synthesis of
Both (Fe = O)+2 (Figure 2) and (Fe = O)+3 ions (Figure 3) abstract H• from the weakest C-H bond present in a molecule to form ferric (Fe+3OH) or ferryl hydroxide (Fe+4OH) and a C• radical respectively [85].
The electrophilic ferric and ferryl hydroxides react ‘instantaneously’ with the nucleophilic C• radical, but the resulting intermediates are different. Ferric hydroxide donates HO• to the C• radical, regenerating the ferrous ion, ending the cycle [33], however the ferryl atom attacks the C• radical (ejecting the hydroxyl group) to form the ferryl-carbon (Fe+4-C) intermediate [83]. Oxygen (O2) insertion into the (Fe+4-C) bond creates the bifurcated oxidative pathways not available to either ferryl-oxo ion or hydroxyl radical [86].
Sugimoto et al. (1987), using 2H and 18O labeled ethanediols, determined that H• abstraction by (Fe = O)+3 from the hydroxyl oxygen of a diol group [R1-HC(O
The Fenton reaction (Fe+2 + H2O2 + H2O) has been shown to generate three powerful oxidants: 1) (HO•) radical [16, 17]; 2) (Fe = O)+2 ion [18]; and 3) [Fe = O]+3 ion [86, 88].
Sugimoto & Sawyer (1985a & 1985b) proposed that both ferrous and ferric ions can abstract an •O• atom from H2O2, thus explaining how ferrous and ferric spontaneously reorganize to form the secondary oxidants ferryl (Fe = O)+2 and perferryl (Fe = O)+3 ions, respectively.
Sugimoto and Sawyer (1984) and (1985b) compared (Fe = O)+2 and (Fe = O)+3 oxidations, respectively, in anhydrous CH3CN or 90% CH3CN/10% H2O with several organic and inorganic molecules. In anhydrous CH3CN, ferryl-oxo ions oxidation produced only 2-electron oxidations, primarily dehydrations or hydroxyl additions, while perferryl-oxo ions produced both 2-, and 4- electron oxidations. Neither oxidant produced 1- electron oxidations.
In aqueous acetonitrile (CH3CN/H2O), single electron oxidations, characteristic of HO• were observed including: 1) carbon–carbon fusions; 2) oxidation of Fe+2 ions; and 3) reduction of Fe+3 ions to Fe+2 ions. The authors proposed that HO• radicals are created by ferryl-oxo and perferryl-oxo ions only when water is present, implying that H• abstraction from water produces HO• radicals via the formula: (Fe = O)+2,+3 + H2O• HO• + Fe(+3,+4)OH [59, 61].
Sawyer et al. (1993) tested the oxidizing capability of Fe+2 ions and organic peroxides (R-O-O-H) 1) under anhydrous conditions in the presence vs. absence of O2, and 2) under anoxic conditions with anhydrous (Fe+2) or partially hydrated (Fe+2(H2O)2) conditions. The authors found evidence of 1e― oxidations either when O2 or H2O2 were present, indicating 1) that (Fe = O)+2 reacted with H2O to form HO• radicals, or 2) with O2, creating O2• [16], which then reacted with (R-O-O-H) to generate HO• radicals [39]. On the other hand, Hage et al. (1995) found that in the conversion of benzene to phenol, if a small amount of H2O was added, the efficiency of conversion was increased, but other 1e― signature products were not detected [89].
Sawyer et al. (1996) surveyed the oxidizing abilities of Fe+2,+3, Cu+2, Co+2, and Mn+2 ions in anhydrous solvents with ROOH, with/out O2. Under an argon atmosphere, only the hydroxyl radical sources produced chain fusion events, none of the ions did. When air (20% O2) was substituted, all of the ions showed oxidation patterns consistent with HO• radicals. The authors concluded that the metal ions, activated by peroxide, reacted with solubilized O2, producing superoxide (O2•− or HO2•), which in turn reacted with H2O2 to generate reactive singlet oxygen (•O•) which then reacts with R-C-H to produce HO• radicals [41].
Barton et al. (1995, 1996) seconded the research of Sawyer’s group, confirming that absent H2O, ferryl-oxo and perferryl-oxo ions perform distinct and distinctive 2- (and 4-) electron oxidations without mixing the unique chemistries of either ion [86, 90].
Mwebi (2005) also confirmed that when Fe+2 ions, H2O2, and H2O are reacted in aqueous conditions, all three secondary oxidants [(Fe = O)+2, (Fe = O)+3, and HO•] arise in that either (Fe = O)+2 and (Fe = O)+3 ions can abstract H• from H2O to create the HO• radical, the HO• radical can oxidize Fe+2 ions to Fe+3 ions, and H2O2 can reduce Fe+3 ions to Fe+2 ions [51].
Oceans covered Earth 4.4 billion years ago [91], evidence of bacteria dates back 3.5 billion years ago (92), and evidence of oxygenic photosynthesis 2.3 billion years ago [91, 92]. From at least that time living organisms have evolved to defend against, and/or, utilize Fenton chemistry.
The use of the Fenton reagents to kill organisms or degrade biopolymers is widely distributed in the biosphere. Saprophytic fungi use Fenton reagents to degrade polysaccharides of woody plant tissues [93], including cellulose [93, 94, 95, 96], callose [97], and hemicelluloses [98].
On the other side of the eukaryote kingdom, mammalian leukocytes and neutrophils pump Fe+2 ions [99, 100] and H2O2 into phagosomes to produce oxygen radicals [101] to effect pathogen killing [102, 103, 104, 105, 106, 107]. For both nutrient mobilization and pathogen killing, these oxidants target external glycan including cell walls to cause cell lysis and/or internal glycans such DNA and RNA to facilitate death of bacteria and eukaryote parasites.
Moore and coworkers incubated
Following Moore’s lead, Lipke and coworkers treated 35S -labeled
In Lim et al. (1995), the authors noted that pretreatment with yeast cells with an Fe+2-binding anti-cancer agent increased cell lysis rates by Zymolyase protease with: 1) Fe+2 + O2 or Fe+3 + O2, but not Ca+2, Co+2, Cu+2, Mn+2, Mg+2, and Zn+2 ions; 2) H2O2 could substitute for O2; and 3) Fe+2/H2O2 and Fe+3/H2O2 alone also accelerated yeast cell lysis; 4) H2O2 only controls did not accelerate Zymolyase lysis rates [112].
To understand the basis of cell wall weakening by Fe+2/H2O2, Ovalle et al. (2001) elected to separately test pure analogs of carbohydrates and proteins found in yeast wall [113]. Ovalle et al. (2001) assumed that partial oxidation of fungal wall monosaccharides would oxidize hydroxyl groups to aldehydes and/or carboxylic acids and developed a method for separating carbohydrates from 0 to 20 glucan units on polyacrylamide gels. Surveying the available literature of the time, the authors followed the method of Ahrgren et al. (1975) where dextran was preincubated with FeSO4 prior to addition of H2O2 [114].
Ovalle et al. (2001) [113] labeled the aldehyde groups of glucose, maltose, maltotriose and enzymatically digested laminaran with 8- amino, 1-, 3-, 6-, naphthalene trisulfonate (ANTS) and NaCNBH3, by the method of Klock & Starr (1998) [115], to have glucan ladders to estimate degree of polymerization of carbohydrate chains separated by polyacrylamide gel electrophoresis. Ovalle et al. (2001) modified the method Klock & Starr (1998) to visualize carboxylic acids and by quenching aldehydes with NaBH4, then crosslinking ANTS to carboxylic acids with N-hydroxysuccinamide (NHS) and N-ethyl-N-(3-aminopropyl) carbodiimide (EDC) [116]. Ovalle et al. (2001) separately visualized de novo aldehydes and de novo carboxylic acids (after quenching aldehydes with NaBH4) on 10% stacking/ 20% running acrylamide gels.
Ovalle et al. (2020) [117] used the method of Ovalle et al. (2001) to determine if Fe+2/H2O2 would oxidize algal laminaran (d.p. ≈ 50–60 glucose units; 97–99% β1–3 glu / 1–3% β1–6 glu). To optimize metal ion-carbohydrate interactions, FeSO4 was incubated with carbohydrate for 1 min prior to addition of H2O2. The final ratio (glucose monomer: Fe+2: H2O2 = 10:1:1) was chosen to oxidize 10% of glucose monomers and reduce the likelihood of a secondary oxidation of glucose fragments to 1% maximum. Unoxidized laminaran did not enter that stacking gel. NaIO4− oxidized laminaran entered the stacking gel but stopped at the stacking gel/running gel interface. Fenton-oxidized laminaran produced smears, when labeled for either aldehyde or carboxylate groups. Enzyme- (Zymolyase) digested laminaran were used as glucan ladders when labeled for aldehydes or organic acids.
To label glucan fragments so as to be suitable for positive ion mass spectroscopy [118, 119], Ovalle et al. (2020) substituted tert-butyl ester of tyrosine (TBT) for ANTS with no other changes required. Ovalle et al. (2020) observed the elution of TBT-labeled glucan fragments with masses consistent with six classes of TBT-labeled molecules: aldoses, dialdoses, uronic acids, hexosuloses, aldonic acids (unlabeled), and hexulosonic acids (unlabeled) (Figure 4).
Comparison of particles of four molecule classes from Laminaran after Fenton oxidation.
Aldose / dialdose pairs arose from glucose by H• abstraction from an unsubstituted hydroxyl groups at O2, O4, or O6, and were mediated by [Fe = O]+2 ion after Fe+2 ion was bound to a site where it was activated by H2O2. Diol-splitting reactions are consistent with oxidation by both HO• and (Fe = O)+2 oxidants, however the ratios of the aldose / dialdose pairs were uneven, implying bias oxidations, hallmark of the (Fe = O)+2 ion.
Uronic and aldonic acids were produced by ketone addition to a hydroxyl carbon (except at C1). The reaction is consistent with oxidation by Barton’s perferryl-oxo ion.
As ferric ions were not added to the assay, Fe+2 ions must have been oxidized by HO• radicals.
Though present, the biased distribution of fragments excludes HO• radicals as the primary oxidant, HO• radicals are partially credited for non-zero values of infrequent carbohydrate fragments. Thus, Ovalle et al. (2020) observed all three Fenton oxidants directly or indirectly in the aqueous Fe+2/H2O2 oxidation of laminaran.
The Fenton Oxidants (HO•, Fe = O+2, and Fe = O+3) are being investigated as molecular scissors for insertion of reactive functional groups into otherwise inert substrates, such as carbohydrates. Oxidation of hydroxyl groups to carbonyl or carboxylic acid groups will allow them to act as carriers for various molecules with ramification in many sectors.
Neyra et al. (2014, 2015) used a catalytic amount Fe+2 ions to produce HO• radicals from H2O2 to oxidize hydroxyl groups of acetylneuraminic acid monomers (2014) and tetramers (2015) to carbonyl and/or carboxylic groups. The goal of the experiment was to modify the sugars to create anchors for proteins so as to create vaccine adjuvant platforms [120, 121].
Sorokin et al. (2004), using ‘heme’-chelated Fe+3 ions, oxidized glucose monomers in starch fibers at C2 and C3 to produce acid / aldehyde pairs without hydrolyzing the flanking glycosidic bridges. The dual oxidations allow for two independent modifications of the glucose monomers in the starch chain [122, 123, 124].
Ozone (O3) is being considered as an alternative to H2O2. Ozone gas can be activated by UV (O3→O2 + •O•) to produce oxygen radicals, or by reaction with iron ions (Fe+2, Fe+3 + O3→Fe = O+2, Fe = O3+ + O2, thus producing each Fenton oxidant without water as a byproduct.
Pestovsky (2004, 2005, 2006) reacted Fe+2 ion with O3 in aqueous buffer as an alternative method of creating (Fe = O)+2 ion. The signature of HO• radicals: 1 e− oxidations, were not detected for the oxidation of several classes of organic molecules [125, 126, 127].
Bataineh (2015a), and Bataineh et al. (2012, 2015b) compared the oxidation of DMSO with Fe+2 and O3 in aqueous phosphate vs. acetonitrile solvents. In acetonitrile the primary product was DMSO2, an oxygen addition reaction. In buffered H2O, ethane and methylsulfinate were the primary products, indicating fragmentation of DMSO occurred by HO• oxidation [128, 129, 130].
Enami et al. (2014) fired microjets of aqueous FeCl2 into sprays of either O2 or O3/O2 mixed gases. Particles detected by negative ion MS proved that Fe+2 and O3 produces new particles not seen in FeCl2 or FeCl2/O2 sprays [131].
Fenton oxidants are gaining popularity as agents of bioremediation because of their ability to mineralize toxic organic molecules without contamination by ecologically damaging elements (halogens, heavy metal ions, etc.).
Turan-Ertas & Gurol (2002) compared ozone (O3) against Fe+3/H2O2 in the degradation of diethylene glycol [(HO-CH2-CH2)O], a toxic byproduct of the synthesis of ethylene glycol. The authors compared the diethylene glycol oxidation profile by O3 and Fe+3/H2O2. Both procedures were effective in degrading diethylene glycol, however the Fe+3/H2O2 oxidation produced fewer and simpler products [132].
In 1894, John HJH Fenton published his discovery that FeSO4 and H2O2 produced oxidations not mimicked by other methods known at the time.
In 1932 & 1934, Fritz Haber & Joseph Weiss proposed the existence of HO• (hydroxyl radical) and HO2•― (superoxide anion) as the principal oxidants of Fenton’s reaction. Merz and Waters (1947) were among the first to propose that HO• radical oxidizes organic molecules by H• abstraction.
In 1932, William Bray & H. M. Gorin proposed (Fe = O)+2 (ferryl-oxo ion) as the principal oxidant of the Fenton’s reaction. Groves and coworkers proposed (in anhydrous solvents) oxygen rebound phenomena, to explain abstraction of H-, followed by addition of HO- to the same carbon to create of alcohols from alkanes in a single step. The debate raged for decades until umpired by D. T. Sawyer and coworkers.
Though 4e− oxidations by Fe+3/H2O2 were observed by Ruff in 1898, and thereafter for nearly 100 years, Derek H. R. Barton & coworkers proposed the structure of the oxidant as (Fe = O)+3 (perferryl-oxo ion) in 1990, and the bifurcated oxygen addition mechanism in 1992.
Donald T. Sawyer & coworkers investigated the behaviors of several transition metal ions in aqueous and anhydrous systems. Sawyer and coworkers proved: 1) H2O is not a spectator molecule; in the absence of water, Fe+2 and Fe+3 ions do not produce HO• radicals, thus explaining why Groves saw only 2e― oxidations in anhydrous media, while Rush and others observed 1e― oxidations in aqueous systems; 2) in aqueous system HO• radicals can oxidize ferrous ions to ferric ions; and 3) HO2•― radicals can reduce ferric ions to ferrous ions, thus in water all three oxidants are present.
Though each oxidant has a singular profile seen in the oxidation of hydrocarbons, different oxidative behaviors are seen with organic molecules containing oxygen. Oxygen causes (Fe = O)+2 and (Fe = O)+3 ions to target hydrogens that are bonded to hydroxyl carbons and hydroxyl oxygens. Abstraction of H• from an O-H bond in molecules with adjacent hydroxyl groups causes C-C cleavage of diols for all three oxidants.
Because of competition between the oxidants for targets, the order of addition of reagents alters the outcome of the assay. Fenton’s method was to begin with substrate, add H2O2, and then FeSO4. In this sequence, when Fe+2 (or Fe+3) ions are activated by peroxide, ferryl-oxo (or perferryl-oxo) ions will react with adjacent H2O molecules, producing HO• radicals, that then diffuse to the substrate, oxidizing H-X bonds by accessibility.
Addition of Fe+2 (or Fe+3) ions first allows the metal ions to associate with and/or chelate onto the substrate. Addition of H2O2 now creates the ferryl-oxo (or perferryl-oxo) ions adjacent to the substrate, increasing the likelihood of in situ oxidation at the ion’s binding site, creating uneven product profiles, as observed in Ovalle et al. (2020). Allowing binding of metal ions to substrates before addition of H2O2 can explain observations of non-canonical “crypto-hydroxyl-” substrate oxidations previously observed by many authors.
This summary is not an exhaustive history, nor is it the full collection of all the articles I read. However, it took me many years to both acquire and understand the chemistry of each oxidant. I did not address other metal/peroxide systems (such as copper-Fenton chemistry) here as it was not relevant to either Ovalle et al. (2001) nor (2020).
This article is written as a guide for newcomers so that they have a head start in finding the papers they need for their research. Welcome to the club!
John H. J. H. Fenton did not know that his discovery would enthrall a legion of researchers, be championed by two Noble laureates, and create three separate fields of peroxide oxidation chemistry: hydroxyl radicals (HO•), ferryl-oxo ions (Fe = O)+2 and perferryl-oxo ions (Fe = O)+3.
Fenton’s successors required a full century to explain the ramification of these reactants. These three simple molecules continue to generate novel research investigations in chemistry, physics, and biology. I am proud to be among Fenton’s successors.
I thank Professors Peter Lipke and Carol Moore for their insights and direction that led me to investigate Fenton chemistry and write Ovalle et al. (2001) and (2020), and Professor Clifford E Soll, who invested hundreds of hours devising the gradient and conditions for sharp separations of the TBT-labeled carbohydrate fragments, but did not live to see the fruition of our collaboration.
Special thanks to Mr. Lijie Chen who acted as my secretary, sounding board, and webmaster as I wrote Ovalle et al. (2020), helping me unearth 100+ years of Fenton literature including the personal histories of the many scientists that followed the path discovered by Prof. Fenton.
I also thank Professors Claude Brathwaite and Barbara Studamire for financial and moral support during the long writing of Ovalle et al. (2020). Finally, I thank Professors Rick Magliozzo, Richard Burger, and Alex Greer for insights into iron-oxygen interactions.
The works of Ovalle et al. (2001) and Ovalle et al. (2020) were supported by NIH GM47176 and RCMI RR03037 grants.
I declare I have no financial or other interests, aside from the telling of the history of, and unusual chemistry of the Fenton reaction and its investigators.
Food emulsions are produced from two immiscible liquids (usually oil and water), which in the presence of an emulsifier and by applying an emulsification method, can be dispersed one into another. Some typical examples include mayonnaise, salad dressings, sauces, milk, ice cream, and sausages. These systems can be used to encapsulate, protect, and deliver biocompounds, including vitamins, flavors, colorants, and nutraceuticals [1]. Emulsifiers are food additives acting by forming a physical barrier between the oil and water, enabling their compatibilization. Effective emulsifiers must be quickly adsorbed at the oil–water interfaces leading to a rapid decrease in the interfacial tension, preventing droplets aggregation. Moreover, they must generate strong repulsive interactions promoting emulsion stability [2, 3].
Synthetic emulsifiers (e.g., Tweens and Spans) are well-known for their ability to form highly stable emulsions. However, consumers’ preferences for healthy, sustainable and natural lifestyle habits have increased worldwide. Moreover, some studies have reported intestinal dysfunctions caused by synthetic emulsifiers [4, 5]. In this context, natural emulsifiers have emerged as great alternatives to replace their conventional counterparts, namely proteins [6], polysaccharides [7], phospholipids [8] and saponins [9]. Concerning protein-based natural emulsifiers, the most use ones come from animal sources (e.g., whey proteins, caseins, egg protein, gelatin) [10]. However, plant-based proteins have demonstrated to be good alternatives for their replacement in products with dietary restrictions (e.g., lactose-free) and in vegetarian and vegan foods. Moreover, plant-based proteins are more sustainable as they have a lower carbon footprint [11, 12]. Examples include pea [13, 14] and soy proteins [15], which have been reported for emulsions production.
Aligned with natural emulsifiers, Pickering stabilizers (in particular organic-based colloidal particles) are emerging as promising solutions. Pickering emulsions or particle-stabilized emulsions present high resistance to coalescence and Oswald ripening due to the tight fixation of the particles to the droplets surface [16]. Several food-grade particles have been studied, namely particles based on proteins [17], polysaccharides [18], and protein/polysaccharide complexes [19]. Furthermore, natural emulsifiers from microbial origin such as biosurfactants and bioemulsifiers are also potential alternatives to be explored in food emulsions [20, 21].
This chapter covers a bibliographic review focused on the last 10-years on natural emulsifiers and emulsion technology field. Research and market trends are also highlighted, showing the most relevant natural emulsifier families. Basic concepts concerning emulsion production, classification, and stabilization methods are introduced. A special emphasis is given to Pickering emulsions regarding novel trends in food emulsion systems.
According to the Research and Markets report, amidst the Covid-19 crisis, the global emulsifiers’ market is projected to reach US$ 6.1 Billion by 2027, growing at a Compound Annual Growth Rate (CAGR) of 4.8% over the forecast period (2020–2027). Particularly, natural emulsifiers’ area is estimated to get US$ 3.3 Billion, recording a 5.4% CAGR [22]. In agreement, the “Global Food Emulsifiers Market 2020-2027 report” from MarketResearch, foresees a high potential for the plant-based emulsifiers in the global food emulsifiers market [23].
Concurrently, scientific literature corroborates the global food emulsifiers report’s projections. More than 8,000 documents were found using the terms “natural emulsifier*” OR “bioemulsifier*” OR “bio-emulsifier*” OR “biosurfactant*” OR “bio-surfactant*” OR “Pickering emulsion*” searched in title, abstract, keywords and Keywords plus sections using the Web of Science Core Collection (SCI-EXPANDED), in the 2010–2020 period. Excluding documents with early publication and applying the “Food Science and Technology” filter from WOS, 792 documents were found. By removing 4 documents from 2021 in a final manual screening, 788 documents were analyzed using Biblioshiny app from the Bibliometrix-R package (RStudio) [24] and VosViewer software [25]. The survey was performed on April 25th, 2021.
Table 1 presents some of the retrieved 788 documents concerning the application of natural emulsifiers or Pickering stabilizers in emulsion formation/stability, including their use in biocompound delivery systems. Some works regarding the production of bioemulsifiers or biosurfactants by microorganisms were also found [31, 32]. Several studies addressing Pickering emulsions and the use of high-pressure homogenization were identified.
Natural emulsifiers or Pickering stabilizers | Emulsification method | Main target | Reference |
---|---|---|---|
Zein-Chitosan complex particles | High-shear homogenization | Delivery system (Curcumin) | [26] |
Mannoprotein | High-shear homogenization | Formation/Stability | [27] |
Whey protein Gum arabic Lecithin | Dual-channel microfluidization | Formation/Stability | [28] |
Gum arabic Beet pectin Corn fiber gum | High-shear homogenization Microfluidizer | Formation/Stability | [7] |
Ginseng saponins | High-shear/ High-pressure homogenization | Delivery system (Astaxanthin) | [9] |
Wheat gluten nanoparticles (WPN) WPN-xanthan gum nanoparticles | High-shear homogenization | Delivery system (β-carotene) | [29] |
Pea protein microgel particles | High-pressure homogenization | Formation/Stability | [30] |
Studies reporting the use of natural molecules and Pickering stabilizers selected from the retrieved 788 documents of the bibliometric search.
Figure 1a shows the wordcloud from Author’s Keyword. The higher font size indicates an increased frequency of the keyword. Figure 1b also illustrates keyword co-occurrence network analysis; the terms distributed in the same cluster present the higher similarity, in comparison with the terms distributed in different clusters.
(a) Wordcloud from Author’s keywords (100 keywords; minimum frequency of 5); (b) keyword co-occurrence network (9 clusters; Author’s keywords; number of occurrences 5).
“Pickering emulsions” is the most frequent keyword, followed by biosurfactant (Figure 1). Other keywords (e.g., whey protein, sodium caseinate, glycolipid, sophorolipids, rhamnolipids,
These findings substantiate the keyword co-occurrence analysis (Figure 1). 93 keywords (Author’s keywords) were organized in 9 clusters. The number of occurrences indicates the number of documents where the keyword appears. Each circle represents a keyword with at least 5 occurrences, being their areas proportional to the number of occurrences. The clusters are characterized by different colors and their words can be related.
Some clusters present words associated to recent trends in the area of natural emulsifiers. Clusters 1, 6, 8 and 9 refer to “Pickering emulsions” and other inter-related words, including nanoparticles, Pickering stabilization, and some commonly used Pickering stabilizers such as starch granules, cellulose nanocrystals and kafirin. Clusters 1 and 2 comprise terms related to the rheological properties of emulsions, an important parameter in food applications. The words included in clusters 4 and 5 are associated with microorganisms (e.g.,
Cluster 7 and 9 are centered in words related to the biocompounds delivery systems, namely bioavailability/bioaccessibility, controlled release, encapsulation and examples of used biocompounds, such as beta-carotene, curcumin, and vitamin E. Clusters 8 and 9 refer to proteins, phospholipids, saponins and polysaccharides, such as whey protein isolate, soy lecithin,
In a general overview, the analysis showed the progressive interest in natural emulsifiers due to their relevance for the scientific and industrial communities, as well as for the global market. Moreover, Pickering emulsions are emerging as advanced emulsion technologies within future trends in the food industry.
Natural emulsifiers belong to a broad range of chemical families and some main examples are shown in Figure 2. Within each family, aspects such as the used natural source or extraction method can lead to different properties. Therefore, the next sections summarize the most relevant families in the area of natural emulsifiers and their contextualization in the field of food applications.
Representative chemical structures for each emulsifier family.
Phospholipids are amphiphilic molecules, and a main constituent of natural membranes. Their structure comprises a hydrophilic head holding a phosphoric acid (H3PO4), combined with a hydrophobic tail composed by one or two non-polar fatty acids. They comprise groups as glycerophospholipids or sphingolipids, with lecithins (glycerophospholipid) assuming an important role. Phospholipids can be obtained from diverse natural sources, including milk, vegetable oils (soybean, rapeseed or sunflower), egg yolk, meat and fish [36, 37]. Specifically, lecithins are known to be good stabilizers for food emulsions, for example the ones derived from soy or egg yolk are applied in mayonnaise, creams, or sauces [38]. Other phospholipid examples include phosphatidylcholine, phosphatidylglycerol, phosphatidylethanolamine, phosphatidylinositol, phosphatidylserine, phosphatidic acid, sphingomyelin. The amphiphilic character of these compounds supports their capacity to stabilize emulsions. Concurrently to their ability to stabilize emulsions they can act as texturizing agents, thus influencing the organoleptic attributes of the final product [39].
Saponins are a complex family derived from plants, constituted by triterpenes or steroid aglycones linked to glycosyl derived sugar structures [40]. Usually the aglycones involve pentacyclic triterpenoids with oleanolic acid and the sugars moieties comprise rhamnose, xylose, glucose or galactose [41]. Factors conditioning the composition of saponins are their botanical origin and extraction method.
Proteins are molecules resulting from the combination of 21 different amino acids, having diverse properties, including water solubility, which varies depending on their composition [46]. Structurally, the presence of both hydrophobic and hydrophilic amino acids confer an amphiphilic character, allowing them to be absorbed at oil/water interfaces, leading to emulsion stabilization [47]. However, proteins have low surface activity in comparison with conventional emulsifiers. This is attributed to the random distribution of the hydrophilic and hydrophobic groups within the peptide chains, limiting their adsorption. This effect is balanced by the protein film formation around the droplets, leading to stabilization through molecular interactions [48]. Diverse proteins (e.g., whey, casein, soy or faba bean proteins) have been tested in food applications, e.g., emulsions for the controlled release of lutein [49], ѡ-3 oil [50], bioactive hydrophobic compounds [51], fish oil [52, 53] or β-carotene [54]. Their application in final products is still hindered by environmental conditions such as pH, temperature and ionic strength [48]. However, these drawbacks can be surpassed by using more complex formulations, namely by combining proteins with polysaccharides [48] or by chemically modifying the proteins trough grafting with other compounds such as polyphenols [54].
Polysaccharides are biopolymers composed of monosaccharide units such as glucose, fructose, mannose or galactose, bonded by glycosidic bonds. Their structural rearrangement, i.e., type and number of monosaccharides, type of glycosidic bonds, molecular weight, electrical charge, branching degree, hydrophobicity and the presence of other groups (carboxylate, sulfate or phosphate), rule the polysaccharides functional properties such as solubility, rheology, and amphiphilic character, among others [10]. Their amphiphilicity depends on the presence of hydrophobic (glycolipids) and hydrophilic (hydroxyls) groups, being adsorbed at the interface, forming a thick stabilizing layer (e.g., pectins, gum Arabic) [55]. Moreover, non-amphiphilic polysaccharides can contribute to emulsion stabilization due to their thickener role, increasing the viscosity and decreasing oil droplets’ motion (e.g., alginates, carrageenan) [56]. Despite the high number of polysaccharides available in nature, only few are authorized as food emulsifiers in EU, namely alginic acid (E400), gum Arabic (E414), pectin (E440), cellulose and chemically modified celluloses (E460 to E469) [57]. Polysaccharides can be obtained from animal, vegetal, microbial fermentation or marine sources (algae), being their properties mostly dependent on the source and extraction process [10].
Microbial synthetic routes are emerging as valuable sustainable and green alternatives to produce emulsifiers. They generate compounds with low ecotoxicity, biodegradability, stability (pH and salinity) and low critical micellar concentration (CMC), in addition to biological activity, biocompatibility and digestibility [58]. Emulsifiers produced by microorganisms are classified according to their molecular weight. Low molecular weight family includes glycolipids (e.g., rhamnolipids, sophorolipids, trehalose lipids) and lipopeptides (e.g., surfactin, iturin, fengycin) and are referred as biosurfactants. Polysaccharides, proteins, lipoproteins, and lipopolysaccharides belong to the high molecular weight family and are referred as bioemulsifiers [59, 60]. Glycolipids like rhamnolipids and trehalose lipids are mostly produced by bacterial strains like
Emulsions are colloidal systems constituted by two immiscible liquids (oil and water), formed in the presence of an emulsifier, and, usually, by applying an energy input. The emulsifier selection is therefore an important step to reach stability. They can be classified based on the hydrophilic region that correspond to ionic structures (anionic or cationic surfactants), change charge with pH (amphoteric surfactants) or present no charged centers (nonionic surfactants) [69]. Among them, nonionic surfactants are often used in food applications because they are less toxic and less affected by pH and ionic strength changes [70, 71]. The choice of a nonionic surfactant can be based on the hydrophilic–lipophilic balance (HLB) index [72]. This scale (0–20), reflects the changing from hydrophobic to hydrophilic character, that is, a lower HLB value corresponds to a lipophilic surfactant being appropriate to stabilize water-in-oil (W/O) emulsions, whereas a high HLB indicates the ability to stabilize oil-in-water (O/W) emulsions, due to the strong hydrophilic balance [72].
Emulsions can be classified according to their typology and structure. The first refers to the relative distribution of the immiscible phases (oil and water), and the latter refers to the arrangement of the emulsified entities [73]. Considering the typology, they can be classified as simple (O/W and W/O) or double (oil-in-water-in-oil (O/W/O), and water-in-oil-in-water (W/O/W)) emulsions (Figure 3). Examples of O/W emulsions in food systems include products such as milk, sauces, beverages, yogurts, ice-creams, and mayonnaise [74]. W/O emulsions are not so frequent but can be found in butter and margarine [73, 75]. For double emulsions, W/O/W are the most used systems due to their ability to generate reduced-fat products, when compared to O/W emulsions. Moreover, they can serve as base systems to encapsulate and control the release of sensitive water-soluble compounds, such as flavors or bioactive ingredients [16, 75, 76].
Typology of simple and double emulsions.
Regarding structure, emulsions can be classified as macroemulsions (usually called emulsions), nanoemulsions, or microemulsions. These systems present specific physicochemical properties that influence their range of applications [71]. Emulsions and nanoemulsions are thermodynamically unstable systems because their free energy is higher than the one of the individual phases [74, 77]. Thus, considering that all systems tend to their lowest energy state, phase separation will occur. However, due to their kinetic stability, they may remain in a metastable state for a considerable period of time, delaying the phase separation phenomenon. The kinetic stability is governed by two mechanisms, namely the energy barriers between the two states (emulsified and separated phases) and mass transfer between the phases. Therefore, high energy barriers and slow mass transfer processes delay phase separation [78]. By contrast, microemulsions are thermodynamically stable systems because their free energy is lower than the one of separate phases. Thus, they can be formed spontaneously under particular compositions and temperature conditions. In practice, some energy input is needed due to the existence of kinetic energy barriers [71]. Regarding the droplet size, nanoemulsions and microemulsions present droplet sizes <200 nm, whereas emulsions hold sizes between 200 nm and 100 μm [16, 71].
Nanoemulsions and microemulsions are optically transparent or slightly turbid due to their small droplet size, being valuable for applications requiring transparency, such as soft drinks [79]. Comparatively with nanoemulsions, microemulsions require a higher emulsifier content, have a lower particle size, and droplets can assume a non-spherical shape, feature that can be used to differentiate the two systems. Emulsions are typically turbid to opaque and are used in creamy systems such as dairy products [80]. Table 2 provides some application examples for each system addressing natural emulsifiers.
System | Type | Natural emulsifiers | Final applications | Reference |
---|---|---|---|---|
Emulsions | O/W | Whey protein | Ice-cream | [81] |
O/W | Coffee creamers | [82] | ||
W/O/W | Whey, rice and pumpkin seed proteins | Cheese | [83] | |
O/W | Faba bean protein | Tofu and yogurts | [84] | |
O/W | Pectin | Functional foods | [85] | |
O/W | Rhamnolipid | Beverages | [86] | |
O/W | Whey protein, locust bean gum, and iota-carrageenan | Mayonnaise, salad dressings, and sauces | [87] | |
O/W | Modified starch | Dairy products | [88] | |
Nanoemulsions | O/W | Soy lecithin | Fruit juices | [89] |
O/W | Soft foods, creams, sauces, and salad dressings | [90] | ||
O/W | Functional foods and beverages | [91] | ||
O/W | Ginseng saponin | [9] | ||
O/W | Whey protein, gum Arabic, and soy lecithin | [92] | ||
O/W | Modified starch, whey and casein proteins | [93] | ||
Microemulsions | W/O | Soy lecithin | Extraction of edible vegetable oils | [94] |
W/O | Soy lecithin | Functional foods and soft drinks | [95] | |
O/W | Soy lecithin | [96] | ||
O/W | Soft drinks and minced meat | [97] |
Food applications of emulsions, nanoemulsions and microemulsions using natural emulsifiers.
Emulsions are thermodynamically unstable mixtures, characterized by the presence of at least two immiscible phases and an emulsifier that, when provided with enough mixing energy, are able to maintain stability over time [98]. The role of the emulsifier is essential to assure stable long-term properties. In general, emulsifiers are active surface substances, enabling their positioning at the oil–water interface, reducing the interfacial tension, hindering (or delaying) aggregation phenomena [99]. Typically, the hydrophilic part of the emulsifier is located in the aqueous phase, while the hydrophobic tail remains enclosed in the oil phase [82, 100]. During emulsion formation, the surfactant molecules require time to move to the interface, forming a layer to reach the interfacial tension equilibrium, a phenomenon related with their adsorption kinetics [82]. This pattern is dependent on emulsifiers’ nature, taking from minutes (e.g., some saponins) to hours (e.g., some proteins), besides being dependent on environmental conditions (e.g., pH, temperature) [82]. To note that, even emulsions are commonly stabilized by a monolayer structure around the droplets, multilayer structures can also be formed. The multilayer pattern favors the electrostatic and steric repulsion of the droplets, improving stability while providing additional protection to the internal phase [16].
The emulsion stabilization mechanism can differ depending on the nature of the used surfactant. In this context four principal stabilization mechanisms are known, namely electrostatic repulsion, steric repulsion, Marangoni-Gibbs effect, and thin film stabilization mechanisms [101]. The electrostatic repulsion is related to ionic emulsifiers and consists on the formation of an electrical double layer at the droplet’s interface, hindering their approximation. Steric repulsion is characteristic of nonionic and/or polymeric emulsifiers, and droplet’s distance is kept due to the adsorption of the hydrophobic segment by the oil phase [101]. The Marangoni-Gibbs effect preserve emulsions’ structure through the deformation of adjacent droplet’s surface, avoiding their outflow, whereas the thin film stabilization mechanism avail the stability of the emulsion by generating a rigid and viscoelastic film, preventing droplets from destabilization effects [101].
Other factors can condition emulsion’s stabilization mechanism, including the emulsifier content, the oil to water ratio or the preparation conditions (pH or temperature). For example, some phospholipids can have no charge at neutral pH, turning into anionic at acidic media, promoting molecule’s swelling at the interface [100]. Moreover, the surfactant concentration can have also impact, e.g., sunflower lecithin in O/W emulsions, at low contents, create a layer surrounding the oil droplets, while at higher concentrations, the stabilization mechanism changes, producing, concurrently, liposomes that might destabilize the emulsion [10]. Considering amphiphilic polymers, when they are used as emulsifiers, they become positioned at the interface, just like the small molecules, but their ability to create intermolecular interactions can provide additional stabilization effects. Their effect on viscosity can also provide a positive stabilization effect [102]. The high hydrophilicity of most polysaccharides can difficult their emulsifier role, if considering the importance of the emulsifiers’ hydrophilic/lipophilic character to interact with both phases. This constraint can be overcome by either chemical or physical strategies [103]. Namely, the suitability of anchoring hydrophobic groups into the polysaccharide structure can equilibrate the hydrophilic/lipophilic balance, that is the hydrophobisation of emulsifier’s surface. Otherwise, alternative approaches imply the mixture of the polysaccharides with other polymers (co-surfactants) to favor the hydrophilic/lipophilic equilibrium and stabilization role.
Food emulsions can be produced using several methods, classified as low-energy and high-energy processes, as represented schematically in Figure 4. The selection of the most appropriate method and respective equipment is based on the volume to process, characteristics of the initial mixture, emulsion’s physicochemical properties, droplet size, and process costs [104]. In Table 3 a survey of recent works dealing with emulsion production trough different methods and using natural emulsifiers in their pure form or compounded with synthetic emulsifiers is presented. Moreover, their potential to encapsulate bioactives for food industry applications is also described.
Schematic representation of the emulsification process through high- and low-energy methodologies.
Productive method | Emulsion type | Oil phase | Particle diameter | Bioactive compound | Emulsifiers | Reference |
---|---|---|---|---|---|---|
Spontaneous emulsification | O/W | MCT | > 10 μm | — | Sunflower phospholipids | [36] |
Emulsion phase inversion | W/O | MCT and orange oil | > 10 μm | Vitamin E | WPI; SMP; Casein; | [105] |
Phase inversion temperature | O/W | Peppermint oil | < 12 nm | Coenzyme Q10 | Lecithin/Tween 20 | [106] |
High-pressure homogenization | O/W | Paprika oleoresin | <150 nm | — | Soy lecithin; WPC; Gum Arabic | [92] |
Microfluidization | O/W | Fish oil | <150 nm | Omega-3 fatty acids | Sunflower phospholipids | [8] |
Colloid mills | W/O | Rapeseed oil | 4.8 μm | — | WPI; Pectin | [107] |
Ultrasonic homogenizer | O/W | MCT; Palm oil; Soybean oil; Rapeseed oil | 0.5–24.1 μm | — | Soy protein isolate | [6] |
Membrane emulsification | W/O/W | Sunflower oil | 35–320 μm | Magnesium | Starch; Pea protein isolate | [108] |
Microchannel homogenizer | O/W | Soybean oil | 35–47 μm | — | Sodium alginate; Pectin; Gum Arabic; Carboxymethyl cellulose | [109] |
High-speed homogenization | O/W | Soybean oil | 143.5 nm | β-carotene; Eugenol | Lecithin; WPI | [110] |
Studies applying different productive methods using natural emulsifiers or natural/synthetic blends to form emulsions and/or to encapsulate biocompounds for food industry applications.
WPI: Whey protein isolate; WPC: Whey protein concentrate; MCT: medium-chain triglycerides; SMP: Sucrose monopalmitate.
Low-energy methods comprise, spontaneous emulsion, and emulsion phase inversion (e.g., phase inversion composition and phase inversion temperature), which occur due to environmental or composition changes namely temperature, pH, and ionic strength of the formulation [104]. Low-energy approaches are more cost effective than high-energy methods. However, they are limited to certain oils and emulsifiers, requiring also large amounts of surfactants, which is not desirable for many food applications [71]. In the work reported by Komaiko et al. [36], spontaneous emulsification lead to emulsions with large droplet size (>10 μm), comparatively with those produced by high-energy methods (<10 μm). The authors concluded that natural emulsifiers can be used in SE emulsions for applications where fine droplets are not essential (Table 3). By contrast, Mayer et al. [105] concluded that it was not possible to produce nanoemulsions using natural emulsifiers by the emulsion phase inversion method. These limitations imply that even natural-based emulsions can be prepared through low-energy methods, high-energy approaches are needed when natural emulsifiers are used.
High-energy methods generate intensive forces promoting the water and oil phases disruption and their subsequent mixture. High-shear homogenizers are the most used equipment’s for producing emulsions in the food industry. They consist on a rotor-stator or stirrer device able to mix the components at high speeds. Usually, large droplets are produced using this approach (1–10 μm) in comparison to alternative high-energy methods. High-pressure homogenization is also widely used in the food industry, being more effective to reduce the droplet size of a pre-emulsion. Generally, this coarse pre-emulsion is produced by high-shear homogenizers, then subjected to the high-pressure homogenization process. The equipment consists of a high-pressure pump (3–500 MPa) to pass the coarse emulsion through a narrow homogenizing valve, generating intensive disruptive forces (shear and cavitation), breaking down the droplets into smaller ones [80, 81].
Many studies have been conducted using two high-energy sequential methods (high-shear and -pressure homogenizers) to produce emulsions/nanoemulsions with natural emulsifiers [111, 112, 113]. Flores-Andrade and co-workers performed a study with soy lecithin, whey protein concentrate (WPC) and gum Arabic as natural emulsifiers, and paprika oleoresin as the oil phase. The coarse emulsion was produced by a high-speed homogenizer, then treated in a high-pressure homogenizer. O/W nanoemulsions were produced, being WPC more effective to form small droplets (d < 150 nm) than the other tested emulsifiers [92].
Microfluidization is the most effective method for producing emulsions with fine droplets (d < 100 nm). This approach is based on feeding the fluid into the homogenizer, which consists of a mixture chamber with two channels. Intensive disruptive forces are generated when these two fluid streams collide at high speed, breaking larger droplets and intermingling the fluids [3]. As the high-pressure homogenizers, microfluidizers were used after preparing a pre-emulsion by high-shear mixers [42, 114]. Ultrasound technique uses high-intense ultrasonic waves, generating intense shear and pressure gradients. The droplets are disrupted mainly by cavitation and turbulent effects [99, 115].
Currently, high-energy approaches are commonly used in the food industry due to their large-scale production capacity and the possibility to process a wide range of raw ingredients [71]. Although several high-energy emulsification devices are available, high-shear and pressure homogenizers, microfluidizers and ultrasound equipment’s are the most used in the production of natural emulsifiers-based emulsions.
Emulsion stability is an important parameter indicating its ability to resist physicochemical changes over time [116]. For food emulsions, the required stability varies according to the intended final application. For example, short-term stability of minutes to hours, is enough for intermediate food emulsions such as cake batter and ice cream mixtures, while long-term stability is required for long shelf-life products, including mayonnaise and salad dressings [117]. For the latter ones, the development of effective strategies to retard emulsion destabilization implies the identification of the main mechanisms leading to this effect [73].
Emulsion instability can occur due to physical and/or chemical processes. The physical instability is responsible for modifying the emulsion droplets spatial distribution and structure, including gravitational separation (creaming/sedimentation), flocculation, coalescence, and Ostwald ripening phenomena (Figure 5). These effects depend on the emulsion composition and structure, besides the storage conditions, namely temperature variation and mechanical stirring [74, 116]. Moreover, the physical phenomena are interrelated and can influence each other during emulsion storage [77].
Common types of instability phenomena in emulsions.
Gravitational separation is driven by density differences between the droplets and the continuous phase. The droplets are subjected to gravitational forces tending to accumulate in the top (creaming) or in the bottom (sedimentation) of the system. Most edible oils present densities lower than water, favoring creaming in O/W emulsions, whereas sedimentation is usually observed in W/O emulsions [116]. Considering the impact of gravitational forces in the large droplets, the separation usually occurs for emulsions with droplet sizes higher than 100 nm or in a final stage of a sequence of instability phenomena [116]. By contrast, for lower droplet sizes, e.g., nanoemulsions, Brownian motion dominates over gravitational forces. Thus, reducing the droplet size is a suitable strategy to retard gravitational separation, with the emulsifier playing an important role to effectively reduce droplets’ size [2, 74]. Furthermore, the emulsifier’ layers tend to minimize the density difference between the emulsion phases, thus reducing the velocity of gravitational separation. Other strategies include modifying the rheology of the continuous phase or increasing the concentration of the droplets [74, 116].
Ostwald Ripening consists of the increase of the droplets size due to the diffusion of small droplets into larger ones, effect driven by their solubility in the continuous phase. This effect is promoted when the droplet’s size decreases [73], being also influenced by the emulsifiers’ properties. Namely, Ostwald Ripening can be retarded by decreasing the interfacial tension of the phases, favored when small-molecule surfactants are used or when using emulsifiers able to form rigid shell around the droplets. By contrast, emulsifiers prone to solubilize the oil and water phases through the formation of colloidal structures (e.g., micelles) accelerate the Ostwald Ripening [2].
Flocculation and coalescence mechanisms are related to droplets aggregation, effect leading to droplet size increase [74]. In flocculation the association of at least two droplets in an aggregate occurs, whereas in the coalescence, the droplets merge into a larger one [77]. Both phenomena are highly dependent on the selected emulsifier [77, 116], namely their nature and colloidal interactions’ capacity [2].
Pickering emulsions are defined as systems stabilized by solid colloidal particles adsorbed at the oil–water interface in a practically irreversible process, creating a coating around the droplets, either in the form of a single or multiple layer, generating a strong steric barrier providing high stability [118]. In the context of Pickering emulsions, the search for natural-based particles is currently a hot topic to face market demands for novel clean label products (absent of emulsifiers) [119]. Pickering emulsions (Figure 6) are raising high interest in the recent years. They are characterized by a long-term stability and have green connotations due to the absence of conventional emulsifiers. These attributes comply with the recent trends of food industry towards the use of sustainable and healthy technologies [16]. The stability of Pickering emulsions is related with the intrinsic properties of the oil and water phases (e.g., type, oil/water ratio, pH, ionic strength) and of the particle stabilizers (e.g., wettability, particle morphology, size and concentration). Particles presenting a contact angle (θ) below 90° are generally suitable for preparing O/W emulsions, whereas θ values greater than 90° indicate good stabilizers for W/O emulsions. At 90°, the particle is immersed equally in both phases [120].
Schematic representation of a Pickering emulsion putting in evidence the particle stabilizers where θ represent the contact angle.
Regarding natural-based particles, three main typologies of stabilizers can be used, namely nanoparticles, microgels and fibrils. Examples include protein derived stabilizers, namely nanoparticles based on corn zein, and colloidal particles of kafirin and gliadin [118, 121, 122, 123]. Although many polysaccharides have high hydrophilic character, some can include hydrophobic side groups (e.g., beet pectin and modified starch) or even active proteins attached to the surface (e.g., gum Arabic) [120], offering potential to act as Pickering stabilizers. Other polysaccharides widely used to produce Pickering bionanoparticles include chitin, chitosan and cellulose. To overcome particle’s limitations as Pickering stabilizers, the formation of complexes has been also proposed, namely complexes such as polysaccharide-polysaccharide, protein–protein, and polysaccharide-protein [124]. Examples include zein-xanthan [125], and tea water insoluble proteins/κ-carrageenan complexes [126].
In the context of the recent trends in Pickering emulsions, research aiming at finding new biological particles, the use of high internal phase emulsions (HIPPE), and the development of bio-based films from Pickering emulsions are becoming topics of high interest for the development of novel food applications. Table 4 presents an overview of recent works dealing with the preparation of Pickering emulsions based on novel biological particles together with the description of the main results envisaging potential food applications.
Particle materials | Main results | Reference |
---|---|---|
Apple pomace | Smaller particles led to emulsions with smaller droplet size, showing higher stability over time (30 days), in addition to improved physical properties (gel-like samples) and antioxidant activity. | [127] |
Bamboo shoots dietary fiber | The emulsions were stable for 4 weeks avoiding coalescence against pH and ionic strength changes and pasteurization conditions. | [128] |
Chitosan-sodium tripolyphosphate | The emulsions presented good thermal stability, showing potential to be applied as a food delivery system for essential oils. | [129] |
Gliadin-pectin | The emulsion with higher particles content (2%) showed suitable physical stability for 30 days, elastic-solid characteristics and good thermal stability (20–80°C). | [130] |
Hordein-chitosan | The emulsions exhibited good stability during storage (14 days, oil ratio = 0.5 and 0.6) and physical properties (elastic gel-like network). | [131] |
Pea protein | The emulsions with higher particle content showed stability against coalescence over 3 months. | [30] |
Sago starch nanocrystals | The emulsions were stable with no signs of creaming for over 2 months. | [132] |
Soy protein isolate-anthocyanin | The emulsions were reached a cream index of 17%, presenting stability for 7 weeks. They presented improved oxidative stability and resistance to | [133] |
Soy protein isolate-chitosan | Cream index values were very low, and the emulsion presented good stability to a broad range of ionic strength and mild temperature conditions (4–60°C). | [134] |
Tea protein | Emulsions with gel-like properties were produced, presenting no creaming over 50 days. | [135] |
Zein-corn fiber gum | High oil concentrations (oil ratio = 0.5) led to higher stability and the formation of a gel-like structure. | [136] |
Zein-gum arabic | The emulsion showed a high stability against coalescence and Ostwald ripening during 30 days of storage (oil ratio = 0.7). | [137] |
Zein-pectin | The emulsions maintained excellent physical stability for 1 month. In addition, they demonstrated good performance as delivery systems of essential oils. | [138] |
β-lactoglobulin-gum arabic | The particles provided stability against coalescence and Ostwald ripening for up to 12 weeks, in addition to improve chemical stability. | [139] |
Examples of bionanoparticles as Pickering stabilizers. All the systems are of O/W type.
HIPPEs are characterized by having a high volume fraction of internal phase (generally higher than 74%), together with relatively low particles concentration resulting in an extremely compacted droplet’s structure [140]. HIPPEs are becoming a novel approach of increasing interest in the food industry, since it combines diverse advantages, namely a semi-solid texture with the ability to encapsulate high amounts of bioactive compounds [141]. HIPPEs allow to control the droplet size distribution, manipulate the morphology and rheological properties, generally presenting enhanced stability against physical, chemical and microbiological stresses [142]. They are positioned as extremely promising substitutes for foods such as margarine, mayonnaise or ice creams [143, 144]. For example, Liu et al. studied wheat gluten as stabilizer in a HIPPE to develop a novel mayonnaise substitute [145]. They obtained excellent results concerning texture and sensory attributes when compared with commercial products.
Bio-based films made from hydrophilic particles added with hydrophobic compounds is another emerging approach in the scope of new applications developed from Pickering emulsions [146]. These strategies provide the ability to improve the stability of the base materials (hydrophilic), in addition to facilitate the combination with hydrophobic materials (e.g., waxes, fatty oils and oils) leading to systems with enhanced moisture barrier properties [147].
The wide variety of emulsion-based systems using natural emulsifiers makes their applicability attractive for various products, particularly in the food industry. The nature and function of emulsifiers, and the formed emulsion type (e.g., nano/micro-scale, simple or double character) can tailor appearance, sensorial characteristics, and attractiveness of foods. Among their diverse functions, the increasing use of emulsions as functionality carriers should be highlighted. In fact, recent works have demonstrated their potential and versatility for the encapsulation of flavors, and to protect and deliver specific bioactives in foods or beverages, helping to strengthen nutritional balances, and enabling the production of reduced-fat products. A summary of examples addressing new trends of emulsion-based products with potential in the food industry are included in Table 5, with some highlighs provided next.
Emulsion type | Oil phase | Emulsifier | Highlights | Reference |
---|---|---|---|---|
O/W | Vegetable oil | Orange pulp and peel powders | Base emulsions for food applications | [148] |
O/W | Orange essential oil | Pea protein concentrate and soy protein isolate | Encapsulation of flavors for the food industry (powder form) | [149] |
O/W | Sunflower, soybean, MCT, and orange oils | Crude saponins isolated from onion skin wastes | Food nanoemulsions (stable emulsions, except at acidic pH and high ionic strength) | [40] |
O/W | Paprika oleoresin | Whey protein and gum Arabic, and soy lecithin | Food and beverage systems for the delivery of carotenoids | [92] |
O/W | Hemp seed oil | Food nanoemulsions with enhanced nutritional properties | [150] | |
O/W | Almond, mustard, olive, and soyabean oils | Biosurfactants isolated from | Food emulsions with improved antibacterial capacity | [151] |
O/W | Corn and sunflower oils) / | Biosurfactant isolated from | Food emulsions with promising properties for salad dressings | [63] |
O1/W/O2 | O1 – Sunflower; O2 - palm oil | Primary emulsion - gelatin, xanthan gum; Secondary emulsion - solid fat crystals | Oil encapsulation systems for texturizing reduced-fat agents | [152] |
Applications of natural-based emulsifiers in food industry.
Lopes Francisco et al. [149] reported an emulsifying system with encapsulation potential based on commercial pea and soy proteins. The work involved the encapsulation of an orange essential oil rich in d-limonene using a O/W emulsion followed by spray drying to obtain powder microparticles. It was demonstrated the ability of pea and soy proteins to act as emulsifiers in the encapsulation of orange essential oil, getting a slightly higher efficiency if using soy protein as the natural emulsifier. These promising results can help consolidate a platform aiming at developing new protective systems to encapsulate flavors for foods, complying with the increasing demand from this industrial sector for natural-based systems.
At the nanoscale, Flores-Andrade et al. [92] reported the preparation of O/W nanoemulsions by high-pressure homogenization, using amphiphilic biopolymers to stabilize paprika oleoresin, namely whey protein, gum Arabic, phospholipids, and soy lecithin. The results demonstrated the effective oil encapsulation, preserving carotenoids (e.g., lipophilic colorants) from chemical degradation, besides positioning this strategy as an attractive route to design new protective and delivery carriers for bioactive compounds aimed at food and/or beverage products.
The potential of double emulsions was also demonstrated by Cetinkaya et al. [152] that evidenced the reduction of the saturated fat content in O1/W/O2 emulsions prepared by fat crystallization according to a two-stage process. Firstly, the primary O1/W emulsion was prepared using sunflower oil and xanthan gum and gelatin as emulsifiers, which was then stabilized in a second oil phase (palm oil), resulting in a structured O1/W/O2 system. Microstructure examination revealed that the accumulation of fat crystals at the interface contributed to stabilize the internal water phase containing the encapsulated sunflower phase. These complex structures showed potential to directly encapsulate hydrophobic oils and act as texturizing reduced-fat agents, which might be of particular interest for the edible oils industry.
This chapter presents an up-to-date overview of current trends in natural emulsifiers and their application in emulsion technology directed to food applications. For this purpose, first, the evolution of food emulsifiers’ scenario over the last 10 years was analyzed through the Bibliometrix-R package (RStudio) and VosViewer software. This analysis indicated a clear driving force towards using natural emulsifiers and the re-emerging importance of the Pickering emulsions. These facts are expected to impact the market growth following the prospectus of available market analysis. The six main identified families of natural emulsifiers were phospholipids, saponins, proteins, polysaccharides, emulsifiers from microbial sources and Pickering stabilizers. Some of them already find extensive use in practical food applications. However, others, mainly natural-based emulsifiers from microbial sources and Pickering stabilizers, despite their high potential, are still needing research investment and regulation clarification (e.g., related to the use of nanoparticles and the use of microbial strains classified as pathogenic in foods). From a technological perspective, the main concepts related to the typology, production methods, stabilization mechanisms, and instability phenomena were presented. Highlighting the increasing interest in Pickering emulsions, a summary of the most recent applications of these systems, including the so-called HIPPEs and their advantages in reduced-fat products development, was provided. To conclude, an analysis of current trends in food emulsion-based products was discussed, putting in evidence the emulsions increasing role as delivery systems of bioactives to support innovative fortified foods advances and the increasing interest in systems based on double emulsions, which provide the opportunity to combine bioactives of different nature. Overall, the field of natural-based emulsifiers combined with the new trends in emulsion technology can, hopefully, be the basis of a new generation of healthy and nutritious food products.
CIMO (UIDB/00690/2020) and AL LSRE-LCM (UIDB/50020/2020) funded by FCT/MCTES (PIDDAC). National funding by FCT, P.I., through the institutional scientific employment program-contract for Arantzazu Santamaria-Echart and Isabel P. Fernandes. FCT for the Research grants (SFRH/BD/148281/2019 of Samara C. da Silva, and SFRH/BD/147326/2019 of Stephany C. de Rezende). GreenHealth project (Norte-01-0145-FEDER-000042).
The authors declare no conflict of interest.
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There is a plethora of research carried out regarding professionalism in policing to meet the needs and challenges of the twenty-first century. Considering the recent developments in police education and training, this chapter mainly discusses three newly introduced routes for recruitment and education of police constables under the Policing Education Qualifications Framework (PEQF), namely Police Constable Degree Apprenticeship (PCDA), Degree Holder Entry Programme (DHEP), and Pre-Join Degree (PJD). Higher education institutions (HEIs), in partnership with the police forces, are providing professional qualifications for policing as a graduate level profession. Though they have made remarkable progress in developing police education programmes, they are facing various challenges in implementing the qualification framework. This chapter also explores pedagogical aspects of police education including the effectiveness and contrast between different forms of teaching and learning. While featuring the challenges and prospects of the new police education programmes, this chapter also outlines different aspects of partnership for delivering these professional qualification programmes.",book:{id:"6950",slug:"education-human-rights-and-peace-in-sustainable-development",title:"Education, Human Rights and Peace in Sustainable Development",fullTitle:"Education, Human Rights and Peace in Sustainable Development"},signatures:"M. Mahruf C. Shohel, Gias Uddin, Julian Parker-McLeod and Daniel Silverstone",authors:[{id:"94099",title:"Dr.",name:"M. Mahruf C.",middleName:null,surname:"Shohel",slug:"m.-mahruf-c.-shohel",fullName:"M. Mahruf C. 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Two UNESCO World Heritage sites will be discussed briefly: Otrar and the surrounding oasis, a medieval complex of sites along the Great Silk Route, and Tamgaly, a petroglyph and archaeological reserve. These two UNESCO World Heritage archaeological sites or preserves will be contrasted with the Talgar Iron Age sites (400 BC–100 CE) situated in a rapidly changing landscape due to economic development and infrastructure (pipelines, railways, roads, and housing) about 12–15 km east of the major city of Almaty. The goal of this article is to discuss the complexity of the entangled sectors of cultural and historic preservation, economic development, tourism, and global transnational heritage within the framework of sustainability.",book:{id:"6950",slug:"education-human-rights-and-peace-in-sustainable-development",title:"Education, Human Rights and Peace in Sustainable Development",fullTitle:"Education, Human Rights and Peace in Sustainable Development"},signatures:"Claudia Chang",authors:[{id:"296402",title:"Dr.",name:"Claudia",middleName:null,surname:"Chang",slug:"claudia-chang",fullName:"Claudia Chang"}]},{id:"71206",doi:"10.5772/intechopen.91053",title:"Uprising and Human Rights Abuses in Southern Cameroon-Ambazonia",slug:"uprising-and-human-rights-abuses-in-southern-cameroon-ambazonia",totalDownloads:949,totalCrossrefCites:1,totalDimensionsCites:2,abstract:"In 2016, lawyers, teachers and students in the two Anglophone regions initially led demonstrations and strikes, which eventually involved a wider section of the population. This mobilization was against their marginalization by the Francophone-dominated government in which they were chronically under-represented in all aspects of national life: political appointments and professional training and had been treated as second-class citizens since their reunification. They argued that their vibrant economic and political institutions had been completely erased, and their education and judicial systems had been undermined and degraded. Activists spread videos that show security forces abusing human rights (by suppressing peaceful gatherings, beating, harassing, arresting and killing protesters, burning their houses, schools and hospitals) in order to produce a counter-narrative to the ‘official story’ that main-stream media had been producing. We collected and analyzed 30 videos to better appreciate the human rights abuses. The videos provide information that cannot be provided by other types of data. They are used as ‘proofs of facts’ and they contain much more visual information on bodily movement and acoustic data. The videos show appalling images not just of how French-speaking soldiers tortured Anglophones but also their inability to communicate with them adequately although they share the same country.",book:{id:"6950",slug:"education-human-rights-and-peace-in-sustainable-development",title:"Education, Human Rights and Peace in Sustainable Development",fullTitle:"Education, Human Rights and Peace in Sustainable Development"},signatures:"Nanche Billa Robert",authors:[{id:"285893",title:"Dr.",name:"Nanche Billa",middleName:null,surname:"Robert",slug:"nanche-billa-robert",fullName:"Nanche Billa Robert"}]}],mostDownloadedChaptersLast30Days:[{id:"68136",title:"Globalization of the Cruise Industry: A Tale of Ships Part II - Asia Post 1994",slug:"globalization-of-the-cruise-industry-a-tale-of-ships-part-ii-asia-post-1994",totalDownloads:926,totalCrossrefCites:1,totalDimensionsCites:1,abstract:"Cruising has grown over 7% a year since 1980. Sustained rapid expansion in North America, followed by local expansion in Europe and Asia, has made cruising a global industry, with 365 ships and estimated sales of $37.8 US billion (CIN, 2017). This global development has been fueled by innovation and introduction of market changing resident ships appealing to the mass traveler which were quickly matched by competitors, establishment of industry and port marketing organizations, awareness of cruising as a vacation option, and availability of suitable port and berthing facilities. When these four conditions coexisted the industry experienced rapid growth. Since 1966, the cruise industry has developed from a Miami-centered industry to a global industry centered in North America, Europe, Asia, and Australia/New Zealand. Given the high cost of state-of-the-art ships, their deployment is a good indication of industry’s confidence in market growth. This chapter chronicles the development of the Asian cruise industry from 1994 through 2017. Data from Cruise Industry News Annual Reports (CIN) and Berlitz Complete Guide to Cruising and Cruise Ships (Ward) are examined and conclusions are drawn.",book:{id:"6950",slug:"education-human-rights-and-peace-in-sustainable-development",title:"Education, Human Rights and Peace in Sustainable Development",fullTitle:"Education, Human Rights and Peace in Sustainable Development"},signatures:"Andrew O. Coggins",authors:[{id:"229658",title:"Prof.",name:"Andrew",middleName:null,surname:"Coggins Jr",slug:"andrew-coggins-jr",fullName:"Andrew Coggins Jr"}]},{id:"72435",title:"Police Education in the United Kingdom: Challenges and Future Directions",slug:"police-education-in-the-united-kingdom-challenges-and-future-directions",totalDownloads:1132,totalCrossrefCites:2,totalDimensionsCites:2,abstract:"This chapter outlines the historical development of police education in the United Kingdom, more precisely in England and Wales, and highlights new strategies and planning for the professional development of the police. There is a plethora of research carried out regarding professionalism in policing to meet the needs and challenges of the twenty-first century. Considering the recent developments in police education and training, this chapter mainly discusses three newly introduced routes for recruitment and education of police constables under the Policing Education Qualifications Framework (PEQF), namely Police Constable Degree Apprenticeship (PCDA), Degree Holder Entry Programme (DHEP), and Pre-Join Degree (PJD). Higher education institutions (HEIs), in partnership with the police forces, are providing professional qualifications for policing as a graduate level profession. Though they have made remarkable progress in developing police education programmes, they are facing various challenges in implementing the qualification framework. This chapter also explores pedagogical aspects of police education including the effectiveness and contrast between different forms of teaching and learning. While featuring the challenges and prospects of the new police education programmes, this chapter also outlines different aspects of partnership for delivering these professional qualification programmes.",book:{id:"6950",slug:"education-human-rights-and-peace-in-sustainable-development",title:"Education, Human Rights and Peace in Sustainable Development",fullTitle:"Education, Human Rights and Peace in Sustainable Development"},signatures:"M. Mahruf C. Shohel, Gias Uddin, Julian Parker-McLeod and Daniel Silverstone",authors:[{id:"94099",title:"Dr.",name:"M. Mahruf C.",middleName:null,surname:"Shohel",slug:"m.-mahruf-c.-shohel",fullName:"M. Mahruf C. Shohel"},{id:"319810",title:"Mr.",name:"Gias",middleName:null,surname:"Uddin",slug:"gias-uddin",fullName:"Gias Uddin"},{id:"321004",title:"Dr.",name:"Julian",middleName:null,surname:"Parker-McLeod",slug:"julian-parker-mcleod",fullName:"Julian Parker-McLeod"},{id:"321005",title:"Dr.",name:"Daniel",middleName:null,surname:"Silverstone",slug:"daniel-silverstone",fullName:"Daniel Silverstone"}]},{id:"73702",title:"Approaches to Analysis of Interstate Cooperation",slug:"approaches-to-analysis-of-interstate-cooperation",totalDownloads:643,totalCrossrefCites:0,totalDimensionsCites:0,abstract:"At the present day cultural diplomacy plays a rather important role in the development of international relations and world politics. This concept is receiving increasing attention from various countries, international and non-governmental organizations and other actors. This trend exists due to a number of reasons, such as the desire of states to create a positive image of their country, the expansion of international cooperation, changes in the global and domestic political situation, the protection of national interests, the prevention of conflicts between states, etc. Cultural diplomacy, beyond historical precedents, consists of a relatively new practice of a country’s foreign policy, which has traditionally focused on trade and security and defense issues. It is true that in European countries there are institutions of cultural foreign relations since the beginning of the century, but in the last decade the issues, related to the projection of the international image of countries, have become more important.",book:{id:"6950",slug:"education-human-rights-and-peace-in-sustainable-development",title:"Education, Human Rights and Peace in Sustainable Development",fullTitle:"Education, Human Rights and Peace in Sustainable Development"},signatures:"Alexander Rozanov, Maria Ivanchenko, Alexandra Baranova, Elena N. Antonova, Mikhail Smirnov, Olga Belyaeva, Maria Ilicheva, Ludmila Ilicheva, Maria Krotovskaya, Tatiana Grabovich, Zaru Utekova, Dmitry Medvedev, Natalya Ogneva, Furat Al-Mutairi, Elvira Shishlo, Amina Surpkelova, Irina Kopachevskaya, Irina Sokurova, Yulia Borisova, Fernando Joao, Artyom Pakulskikh, Polina Chernova, Alexandra Khramova, Oksana Gryuk, Jesus Yaniz Gonzalez, Valentina Komleva, Alina Papsheva and Arkadi Bessonov",authors:[{id:"233092",title:"Dr.",name:"Alexander",middleName:null,surname:"Rozanov",slug:"alexander-rozanov",fullName:"Alexander Rozanov"},{id:"312194",title:"Prof.",name:"Valentina",middleName:"Vycheslavovna",surname:"Komleva",slug:"valentina-komleva",fullName:"Valentina Komleva"},{id:"312195",title:"Ms.",name:"Alexandra",middleName:null,surname:"Baranova",slug:"alexandra-baranova",fullName:"Alexandra Baranova"},{id:"312196",title:"Dr.",name:"Furat",middleName:null,surname:"Al Mutairi",slug:"furat-al-mutairi",fullName:"Furat Al Mutairi"},{id:"312197",title:"Ms.",name:"Maria",middleName:null,surname:"Ivanchenko",slug:"maria-ivanchenko",fullName:"Maria Ivanchenko"},{id:"312198",title:"Associate Prof.",name:"Arkadi",middleName:null,surname:"Bessonov",slug:"arkadi-bessonov",fullName:"Arkadi Bessonov"},{id:"312199",title:"Ms.",name:"Alina",middleName:null,surname:"Papsheva",slug:"alina-papsheva",fullName:"Alina Papsheva"},{id:"312200",title:"Prof.",name:"Ludmila",middleName:null,surname:"Ilicheva",slug:"ludmila-ilicheva",fullName:"Ludmila Ilicheva"},{id:"312201",title:"Ph.D. Student",name:"Aleksandra",middleName:null,surname:"Khramova",slug:"aleksandra-khramova",fullName:"Aleksandra Khramova"},{id:"316768",title:"Dr.",name:"Maria",middleName:null,surname:"Ilicheva",slug:"maria-ilicheva",fullName:"Maria Ilicheva"},{id:"317753",title:"Dr.",name:"Oksana",middleName:null,surname:"Gryuk",slug:"oksana-gryuk",fullName:"Oksana Gryuk"}]},{id:"71206",title:"Uprising and Human Rights Abuses in Southern Cameroon-Ambazonia",slug:"uprising-and-human-rights-abuses-in-southern-cameroon-ambazonia",totalDownloads:951,totalCrossrefCites:1,totalDimensionsCites:2,abstract:"In 2016, lawyers, teachers and students in the two Anglophone regions initially led demonstrations and strikes, which eventually involved a wider section of the population. This mobilization was against their marginalization by the Francophone-dominated government in which they were chronically under-represented in all aspects of national life: political appointments and professional training and had been treated as second-class citizens since their reunification. They argued that their vibrant economic and political institutions had been completely erased, and their education and judicial systems had been undermined and degraded. Activists spread videos that show security forces abusing human rights (by suppressing peaceful gatherings, beating, harassing, arresting and killing protesters, burning their houses, schools and hospitals) in order to produce a counter-narrative to the ‘official story’ that main-stream media had been producing. We collected and analyzed 30 videos to better appreciate the human rights abuses. The videos provide information that cannot be provided by other types of data. They are used as ‘proofs of facts’ and they contain much more visual information on bodily movement and acoustic data. The videos show appalling images not just of how French-speaking soldiers tortured Anglophones but also their inability to communicate with them adequately although they share the same country.",book:{id:"6950",slug:"education-human-rights-and-peace-in-sustainable-development",title:"Education, Human Rights and Peace in Sustainable Development",fullTitle:"Education, Human Rights and Peace in Sustainable Development"},signatures:"Nanche Billa Robert",authors:[{id:"285893",title:"Dr.",name:"Nanche Billa",middleName:null,surname:"Robert",slug:"nanche-billa-robert",fullName:"Nanche Billa Robert"}]},{id:"72097",title:"Towards Global Peace and Sustainability: Role of Education in Peace-Building in the Great Lakes Region of Sub-Saharan Africa",slug:"towards-global-peace-and-sustainability-role-of-education-in-peace-building-in-the-great-lakes-regio",totalDownloads:683,totalCrossrefCites:0,totalDimensionsCites:0,abstract:"The Great Lakes Region of sub-Saharan Africa is well known for being volatile and turbulent in terms of peace and stability. For over 60 years, almost all countries in the region have experienced some kind of political and social turmoil such as civil war, coup de tat, and genocides. In 1960, the first democratically elected Congolese prime minister was assassinated. There were unprecedented social and political havoc in a nearby “other Congo” characterized by power struggle between various political and ethnic factions in the post-independence Congo Brazzaville. In Burundi and Rwanda, ethnic tensions between the Tutsi and Hutu engulfed the developmental dreams of nationalist freedom fighters until 2015. Though arguably stable, Tanzania has experienced its own share of socio-political messy including the 1998 Mwembechai and 2001 Pemba massacres. Efforts to build a sense of sustainable peace and development based on mutual understanding and socio-political harmony has brought limited success. In all these countries, the missing link in building sustainable peace and security has been a lack of education. The chapter intends to fill this gap by critically analyzing the potential role of basic education, especially pre-primary and early grades education, in sustainable peace-building in the sub-Saharan context.",book:{id:"6950",slug:"education-human-rights-and-peace-in-sustainable-development",title:"Education, Human Rights and Peace in Sustainable Development",fullTitle:"Education, Human Rights and Peace in Sustainable Development"},signatures:"Laurent Gabriel Ndijuye and Pambas Basil Tandika",authors:[{id:"301740",title:"Dr.",name:"Laurent Gabriel",middleName:null,surname:"Ndijuye",slug:"laurent-gabriel-ndijuye",fullName:"Laurent Gabriel Ndijuye"},{id:"319403",title:"Dr.",name:"Pambas Basilius",middleName:null,surname:"Tandika",slug:"pambas-basilius-tandika",fullName:"Pambas Basilius Tandika"}]}],onlineFirstChaptersFilter:{topicId:"476",limit:6,offset:0},onlineFirstChaptersCollection:[],onlineFirstChaptersTotal:0},preDownload:{success:null,errors:{}},subscriptionForm:{success:null,errors:{}},aboutIntechopen:{},privacyPolicy:{},peerReviewing:{},howOpenAccessPublishingWithIntechopenWorks:{},sponsorshipBooks:{sponsorshipBooks:[],offset:8,limit:8,total:0},allSeries:{pteSeriesList:[{id:"14",title:"Artificial Intelligence",numberOfPublishedBooks:9,numberOfPublishedChapters:90,numberOfOpenTopics:6,numberOfUpcomingTopics:0,issn:"2633-1403",doi:"10.5772/intechopen.79920",isOpenForSubmission:!0},{id:"7",title:"Biomedical Engineering",numberOfPublishedBooks:12,numberOfPublishedChapters:107,numberOfOpenTopics:3,numberOfUpcomingTopics:0,issn:"2631-5343",doi:"10.5772/intechopen.71985",isOpenForSubmission:!0}],lsSeriesList:[{id:"11",title:"Biochemistry",numberOfPublishedBooks:33,numberOfPublishedChapters:330,numberOfOpenTopics:4,numberOfUpcomingTopics:0,issn:"2632-0983",doi:"10.5772/intechopen.72877",isOpenForSubmission:!0},{id:"25",title:"Environmental Sciences",numberOfPublishedBooks:1,numberOfPublishedChapters:19,numberOfOpenTopics:4,numberOfUpcomingTopics:0,issn:"2754-6713",doi:"10.5772/intechopen.100362",isOpenForSubmission:!0},{id:"10",title:"Physiology",numberOfPublishedBooks:14,numberOfPublishedChapters:145,numberOfOpenTopics:4,numberOfUpcomingTopics:0,issn:"2631-8261",doi:"10.5772/intechopen.72796",isOpenForSubmission:!0}],hsSeriesList:[{id:"3",title:"Dentistry",numberOfPublishedBooks:9,numberOfPublishedChapters:139,numberOfOpenTopics:2,numberOfUpcomingTopics:0,issn:"2631-6218",doi:"10.5772/intechopen.71199",isOpenForSubmission:!0},{id:"6",title:"Infectious Diseases",numberOfPublishedBooks:13,numberOfPublishedChapters:122,numberOfOpenTopics:4,numberOfUpcomingTopics:0,issn:"2631-6188",doi:"10.5772/intechopen.71852",isOpenForSubmission:!0},{id:"13",title:"Veterinary Medicine and Science",numberOfPublishedBooks:11,numberOfPublishedChapters:112,numberOfOpenTopics:3,numberOfUpcomingTopics:0,issn:"2632-0517",doi:"10.5772/intechopen.73681",isOpenForSubmission:!0}],sshSeriesList:[{id:"22",title:"Business, Management and Economics",numberOfPublishedBooks:1,numberOfPublishedChapters:21,numberOfOpenTopics:3,numberOfUpcomingTopics:0,issn:"2753-894X",doi:"10.5772/intechopen.100359",isOpenForSubmission:!0},{id:"23",title:"Education and Human Development",numberOfPublishedBooks:0,numberOfPublishedChapters:10,numberOfOpenTopics:1,numberOfUpcomingTopics:1,issn:null,doi:"10.5772/intechopen.100360",isOpenForSubmission:!0},{id:"24",title:"Sustainable Development",numberOfPublishedBooks:1,numberOfPublishedChapters:19,numberOfOpenTopics:5,numberOfUpcomingTopics:0,issn:"2753-6580",doi:"10.5772/intechopen.100361",isOpenForSubmission:!0}],testimonialsList:[{id:"13",text:"The collaboration with and support of the technical staff of IntechOpen is fantastic. The whole process of submitting an article and editing of the submitted article goes extremely smooth and fast, the number of reads and downloads of chapters is high, and the contributions are also frequently cited.",author:{id:"55578",name:"Antonio",surname:"Jurado-Navas",institutionString:null,profilePictureURL:"https://s3.us-east-1.amazonaws.com/intech-files/0030O00002bRisIQAS/Profile_Picture_1626166543950",slug:"antonio-jurado-navas",institution:{id:"720",name:"University of Malaga",country:{id:null,name:"Spain"}}}},{id:"6",text:"It is great to work with the IntechOpen to produce a worthwhile collection of research that also becomes a great educational resource and guide for future research endeavors.",author:{id:"259298",name:"Edward",surname:"Narayan",institutionString:null,profilePictureURL:"https://mts.intechopen.com/storage/users/259298/images/system/259298.jpeg",slug:"edward-narayan",institution:{id:"3",name:"University of Queensland",country:{id:null,name:"Australia"}}}}]},series:{item:{id:"10",title:"Physiology",doi:"10.5772/intechopen.72796",issn:"2631-8261",scope:"Modern physiology requires a comprehensive understanding of the integration of tissues and organs throughout the mammalian body, including the cooperation between structure and function at the cellular and molecular levels governed by gene and protein expression. 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From\r\n1964 to 1974, he worked as Assistant in Biochemistry at the School of MedicineUniversidad Nacional de La Plata, Argentina. From 1974 to 1976, he was a Fellowof the National Institutes of Health (NIH) at the University of Connecticut, Health Center, USA. From 1985 to 2004, he served as a Full Professor oBiochemistry at the Universidad Nacional de La Plata, Argentina. He is Member ofthe National Research Council (CONICET), Argentina, and Argentine Society foBiochemistry and Molecular Biology (SAIB). His laboratory has been interested for manyears in the lipid peroxidation of biological membranes from various tissues and different species. Professor Catalá has directed twelve doctoral theses, publishedover 100 papers in peer reviewed journals, several chapters in books andtwelve edited books. Angel Catalá received awards at the 40th InternationaConference Biochemistry of Lipids 1999: Dijon (France). 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