270 The nutrition handbook for food processors CHO C=N-R CH-NH-R CHOH RNH CHOH reaction CHOH 5 CHOH CH=O CHOH -H2O CHOH CHOH R R Fig. 11.5 Possible pathway for the formation of glycolaldehyde alkylamines proposed by nd Hayashi (1986 R-NH2 Ho CH2OH CH,OH A CH2OH HoH F(Ct +Cs) F(c2+c4) route O(NR) NHR CH2OH CHOH CH2OH cyclisation 3. 4-dideoxy Fig. 11.6 Transformation of hexoses and pentoses to Cs- and Ca-pyrroles and-furans. (Reproduced with permission from Tressel et al, 1998a) However, in the last two decades other mechanisms have been proposed. For example, starting from the experimental observation of free radical formation at the start of the MR, Hayashi and Namiki(1981: 1986) proposed a reducing sugar degradation pathway that produces glycolaldehyde alky limine without passin through the formation of APs(Fig. 11.5 Very recently, on the basis of extensive experiments with- H-labelled ugars,a detailed reaction scheme was proposed by Tressel et al (1995 and 1998a): the formation of various C6", Cs", and Ca-pyrroles and furans from both intact and fragmented hexoses and amines could be unambiguously attributed to distinct reaction pathways via the intermediates A-C without involving the
However, in the last two decades other mechanisms have been proposed. For example, starting from the experimental observation of free radical formation at the start of the MR, Hayashi and Namiki (1981; 1986) proposed a reducing sugar degradation pathway that produces glycolaldehyde alkylimines without passing through the formation of ARPs (Fig. 11.5). Very recently, on the basis of extensive experiments with 13C- and 2 H-labelled sugars, a detailed reaction scheme was proposed by Tressel et al (1995 and 1998a): the formation of various C6-, C5-, and C4-pyrroles and furans from both intact and fragmented hexoses and amines could be unambiguously attributed to distinct reaction pathways via the intermediates A–C without involving the 270 The nutrition handbook for food processors CHO CHOH CHOH CHOH R' C=N-R CHOH CHOH CHOH R' CH-NH-R CHOH CH=O CHOH R' + RNH2 - H2O reverse aldol reaction 5 Fig. 11.5 Possible pathway for the formation of glycolaldehyde alkylimines proposed by Namiki and Hayashi (1986). CHO OH HO OH OH CH2OH OH HO OH OH CH2OH NR O H OH OH CH2OH NR O H OH CH2OH NR O H O OH CH2OH NHR O H OH OH CH2OH O H OH CH2OH O H OH OH CH2OH O H CH2OH NR OH H O NR O H O CH2OH NHR OH H O O NHR N R OH O OH O N R O O O N R polymers polymers 2-deoxypentoses tetroses pentoses C6-pyrroles C6-furans cyclisation cyclisation beta-dicarbonyl route 3,4-dideoxy aldoketose route 3,4-dideoxy aldoketose route 3,4-dideoxy aldoketose route beta-dicarbonyl route F(c1 + c5) F(c2 + c4) F(c5 + c1) F(c5 + c1) R-NH2 3-deoxy aldoketose route (NR) A B C H H H H (NR) Fig. 11.6 Transformation of hexoses and pentoses to C5- and C4-pyrroles and -furans. (Reproduced with permission from Tressel et al, 1998a)
Thermal processing and nutritional quality 271 Table 11.1 Composition of the primary fragmentation pools Type of pool Constituents Amino acid Amines fragmentation pool Carboxylic acids Kanes and aromatics Aldehydes Amino acid specific side chain fragments: H,S(Cys), CH,SH even Sugar fragmentation CI fragments: formaldehyde, formic acid pool (S) C2 fragments: glyoxal, glycoladehyde, acetic acid 3 fragments: glyceraldehyde, methylglyoxal, hydroxyacetone dihydroxyacetone, etc. C4 fragments: tetroses, 2, 3-butanedione, 1-hydroxy-2-butanone 2-hydroxybutanal, etc. C5 fras eoxy derivati C6 fragments: pyranone, furans, glucosone, deoxyglucosone Amadori and Heyns C3-ARP/HRP derivatives: glyceraldehyde-ARP, amino fragmentation pool acid-propanone, amino acid-propanal, etc IDI C4-ARP/HRP derivatives: amino acid-tetradiuloses amino acid-butanones C5-ARP/HRP derivatives: amino acid-pentadiuloses C6-ARP/HRP derivatives: amino acid-hexadiuloses, pyrylium betaines Lipid fragmentation Propanal, al hexanal, octanal. nonanal 2-Oxoaldeh C6-9) C2 fragments: glyoxal 3 fragments: CHOCH_CHO, methylglyoxal Amadori rearrangement(Fig. 11.6). These pyrroles and furans polymerise very easily to highly coloured compounds that may be involved in the formation of melanoidins By means of experiments showing that sugars and most amino acids also undergo independent degradation(Yaylayan and Keyhani, 1996), a new concep tual approach to the Mr has been proposed recently by Yaylayan(1997). He sug- gested that in order to understand the mr better. it is more useful to define a sugar fragmentation pool [S), an amino acid fragmentation pool (A], and an interaction fragmentation pool (D, deriving from the Amadori and Heyns com- pounds( Table 11. 1). Together they constitute a primary fragmentation pool of building blocks that react to give a secondary pool of interaction intermediates and eventually a very complex final pool of stable end-products G owever. most foods contain also lipids that can degrade by autoxidation rosch, 1987) giving reactive intermediates, mainly saturated or unsaturated aldehydes or ketones and also glyoxal and methylglyoxal (in common with the
Amadori rearrangement (Fig. 11.6). These pyrroles and furans polymerise very easily to highly coloured compounds that may be involved in the formation of melanoidins. By means of experiments showing that sugars and most amino acids also undergo independent degradation (Yaylayan and Keyhani, 1996), a new conceptual approach to the MR has been proposed recently by Yaylayan (1997). He suggested that in order to understand the MR better, it is more useful to define a sugar fragmentation pool {S}, an amino acid fragmentation pool {A}, and an interaction fragmentation pool {D}, deriving from the Amadori and Heyns compounds (Table 11.1). Together they constitute a primary fragmentation pool of building blocks that react to give a secondary pool of interaction intermediates and eventually a very complex final pool of stable end-products. However, most foods contain also lipids that can degrade by autoxidation (Grosch, 1987) giving reactive intermediates, mainly saturated or unsaturated aldehydes or ketones and also glyoxal and methylglyoxal (in common with the Thermal processing and nutritional quality 271 Table 11.1 Composition of the primary fragmentation pools Type of pool Constituents Amino acid Amines fragmentation pool Carboxylic acids {A} Alkanes and aromatics Aldehydes Amino acid specific side chain fragments: H2S (Cys), CH3SH (Met), styrene (Phe) Sugar fragmentation C1 fragments: formaldehyde, formic acid pool {S} C2 fragments: glyoxal, glycoladehyde, acetic acid C3 fragments: glyceraldehyde, methylglyoxal, hydroxyacetone, dihydroxyacetone, etc. C4 fragments: tetroses, 2, 3-butanedione, 1-hydroxy-2-butanone, 2-hydroxybutanal, etc. C5 fragments: pentoses, pentuloses, deoxy derivatives, furanones, furans C6 fragments: pyranones, furans, glucosones, deoxyglucosones Amadori and Heyns C3-ARP/HRP derivatives: glyceraldehyde-ARP, amino fragmentation pool acid-propanone, amino acid-propanal, etc. {D} C4-ARP/HRP derivatives: amino acid-tetradiuloses, amino acid-butanones C5-ARP/HRP derivatives: amino acid-pentadiuloses C6-ARP/HRP derivatives: amino acid-hexadiuloses, pyrylium betaines Lipid fragmentation Propanal, pentanal, hexanal, octanal, nonanal pool {L} 2-Oxoaldehydes (C6–9) C2 fragments: glyoxal C3 fragments: CHOCH2CHO, methylglyoxal Formic acid, acids
272 The nutrition handbook for food processors Interaction pool A S heterocycles D}{} Pri fragmentation pool dimers Fig. 11.7 Conceptual representation of the Maillard reaction: genealogy of primary ragmentation pools, interaction pools (containing self-interaction pools as well as mix interaction pools) and end-products Maillard reaction) and malondialdehyde(Table 11.1). These belong to a fourth pool, the lipid fragmentation pool (L)(' Agostina et al, 1998)and in this way the scheme proposed by Yaylayan(1997)was revised to include it(Fig. 11.7) Clear interconnections between the MR and lipid autoxidation have been exten sively studied in the case of food aromas, where many end-products deriving from lipids and amino acids or sugars are very well documented(whitfield, 1992), but certainly they may be relevant also for other sensory aspects, such as colour or taste, or for nutrition, although these research areas have been almost completely neglected until the present. Depending on food composition and heating intensity applied, thousands of fferent end products may be formed in the advanced stage of the MR: they are classified here according to their functions in foods(Fig. 11.8). Very volatile com- pounds, such as pyrazines, pyridines, furans, thiophenes, thiazoles, thiazolines, and dithiazine are of interest when considering aroma: some low molecular weight compounds relate to taste(Frank et al, 2001: Ottiger et al, 2001), others behave as antioxidants and a few are mutagenic. Polymers(melanoidins) that in sugar/amino acid model systems and some foods such as coffee, roasted malt, or chocolate are the major MRPs, and determine the colour of the food. This review will discuss only the mechanism of formation of MRPs that have some nutritional significance or may be used as molecular markers for quantify- ing the MR in foods. A very detailed description of the pathways leading to most Maillard reaction icts may be found in an excellent review by ledl and Schleicher(1990)
Maillard reaction) and malondialdehyde (Table 11.1). These belong to a fourth pool, the lipid fragmentation pool {L} (D’Agostina et al, 1998) and in this way the scheme proposed by Yaylayan (1997) was revised to include it (Fig. 11.7). Clear interconnections between the MR and lipid autoxidation have been extensively studied in the case of food aromas, where many end-products deriving from lipids and amino acids or sugars are very well documented (Whitfield, 1992), but certainly they may be relevant also for other sensory aspects, such as colour or taste, or for nutrition, although these research areas have been almost completely neglected until the present. Depending on food composition and heating intensity applied, thousands of different end products may be formed in the advanced stage of the MR: they are classified here according to their functions in foods (Fig. 11.8). Very volatile compounds, such as pyrazines, pyridines, furans, thiophenes, thiazoles, thiazolines, and dithiazines are of interest, when considering aroma; some low molecular weight compounds relate to taste (Frank et al, 2001; Ottiger et al, 2001), others behave as antioxidants and a few are mutagenic. Polymers (melanoidins) that in sugar/amino acid model systems and some foods such as coffee, roasted malt, or chocolate are the major MRPs, and determine the colour of the food. This review will discuss only the mechanism of formation of MRPs that have some nutritional significance or may be used as molecular markers for quantifying the MR in foods. A very detailed description of the pathways leading to most Maillard reaction products may be found in an excellent review by Ledl and Schleicher (1990). 272 The nutrition handbook for food processors {A} {S} {D} {L} polymers Interaction pool Primary fragmentation pool oligomers dimers heterocycles Fig. 11.7 Conceptual representation of the Maillard reaction: generalogy of primary fragmentation pools, interaction pools (containing self-interaction pools as well as mixinteraction pools) and end-products
Thermal processing and nutritional quality 273 Volatile compounds Antioxidants Maillard reaction Toxic compounds Tasty compounds Brown compound Fig. 11.8 Functional classification of Maillard reaction products 11.3 Nutritional consequences and molecular markers of the mailllard reaction in food As the Mr involves some of the most important food nutrients, its nutritional consequences must be carefully considered. Researcher attention has previously been focused mainly on milk and milk products, where thermal treatments are necessary for obtaining microbial stabilisation and the preservation of high nutritional quality. Vegetable products, which become edible only after thermal treatments, have been relatively neglected so far The degradation of sugars per se is never considered a problem because it is only rarely they are lacking in diet. However, free or protein-bound essential amino acids may be damaged irreversibly; the amount of free amino acids in food is always very low and they are important as constituents of proteins. This means that the most relevant nutritional effect of the Maillard reaction is non-enzymatic glycosylation of proteins which involves mostly lysine, whose bioavailability may be drastically impaired. This should be distinguished very clearly from enzy matic glycosylation, a normal step in the biosynthesis of glycoproteins, in which oligosaccharides are bound to serine or asparagine through a glycosidic bond. The first glycation products are then converted to the Amadori product, fruc tosyllysine, that eventually can cross-link with other amino groups intramolecu larly or intermolecularly. The resulting polymeric aggregates are called advanced glycation end products(AGEs) Lysine availability is an important nutritional parameter especially in foods for particular classes of consumers, such as infant formulas(Ferrer et al, 2000) Statistically significant losses of available lysine(about 20%)with respect to raw milk have been reported as a consequence of the thermal treatment applied in the preparation of these foods Because the reactions of lysine are so relevant in nutrition, over a period of time different MRPs have been proposed as markers of protein glycosylation
11.3 Nutritional consequences and molecular markers of the Maillard reaction in food As the MR involves some of the most important food nutrients, its nutritional consequences must be carefully considered. Researcher attention has previously been focused mainly on milk and milk products, where thermal treatments are necessary for obtaining microbial stabilisation and the preservation of high nutritional quality. Vegetable products, which become edible only after thermal treatments, have been relatively neglected so far. The degradation of sugars per se is never considered a problem because it is only rarely they are lacking in diet. However, free or protein-bound essential amino acids may be damaged irreversibly; the amount of free amino acids in food is always very low and they are important as constituents of proteins. This means that the most relevant nutritional effect of the Maillard reaction is non-enzymatic glycosylation of proteins which involves mostly lysine, whose bioavailability may be drastically impaired. This should be distinguished very clearly from enzymatic glycosylation, a normal step in the biosynthesis of glycoproteins, in which oligosaccharides are bound to serine or asparagine through a glycosidic bond. The first glycation products are then converted to the Amadori product, fructosyllysine, that eventually can cross-link with other amino groups intramolecularly or intermolecularly. The resulting polymeric aggregates are called advanced glycation end products (AGEs). Lysine availability is an important nutritional parameter especially in foods for particular classes of consumers, such as infant formulas (Ferrer et al, 2000). Statistically significant losses of available lysine (about 20%) with respect to raw milk have been reported as a consequence of the thermal treatment applied in the preparation of these foods. Because the reactions of lysine are so relevant in nutrition, over a period of time different MRPs have been proposed as markers of protein glycosylation. Thermal processing and nutritional quality 273 Volatile compounds Maillard reaction Antioxidants Metal chelating agents Toxic compounds Tasty compounds Brown compounds Fig. 11.8 Functional classification of Maillard reaction products
274 The nutrition handbook for food processors H R'OC-C-NHR (CH2)4 lysine (CH2)4 N-(CH2)4 HO→H (CHa) CH2OH H3C、N Fig. 11.9 Compounds derived from fructosyllysine decomposition. Fructosyllysine is unstable in the acid conditions of protein hydrolysis, produc- ing about 30% furosine, pyridosine(a minor cyclisation product) and about 50% lysine(Fig. 11.9). Furosine was first detected in foods by Erbersdobler and Zucker (1966)and can be easily analysed by HPLC: thus furosine quantification is con- sidered a good estimate of nutritionally unavailable lysine Milk proteins, owing to their nutritional relevance, have been considered with particular attention Owing to the presence of lactose, the Amadori compound in this case is lactulo- syllysine and furosine is again a useful marker of lysine unavailability. For this reason several authors have used the furosine method for determining the progress of the Maillard reaction in different foods( Chiang, 1983: Hartkopf and Erbersdobler, 1993 and 1994, Henle et al, 1995; Resmini et al, 1990). However, today very powerful analytical techniques are disclosing new possibilities permitting, for example, the direct determination of fructosyllysine (Vinale et al, 1999) by the use of a stable isotope dilution assay performed in liquid chromatography- mass spectrometry(LC-MS). This method overcomes the problems of hydrolytic instability of the analyte and the incompleteness of the enzymatic digestion technique Other possible markers of lysine transformation are N-E-carboxymethyllysine (CML) and 5-hydroxymethylfurfural (HMF)(Fig. 11.10). CML was detected for the first time in milk by Buser and Erbersdobler (1986)and an oxidative mech- anism was proposed for its formation(Ahmed et al, 1986).The formation of HMF in foods has been explained in two ways: via the Amadori products through eno- lisation (in the presence of amino groups)and via lactose isomerisation and degra- dation, known as the lobry de Bruyn-Alberda van Ekenstein transformation (Ames, 1992). Because of this, it has recently been proposed to measure sepa- rately the HMF formed only by the acidic degradation of Amadori products and
Fructosyllysine is unstable in the acid conditions of protein hydrolysis, producing about 30% furosine, pyridosine (a minor cyclisation product) and about 50% lysine (Fig. 11.9). Furosine was first detected in foods by Erbersdobler and Zucker (1966) and can be easily analysed by HPLC: thus furosine quantification is considered a good estimate of nutritionally unavailable lysine. Milk proteins, owing to their nutritional relevance, have been considered with particular attention. Owing to the presence of lactose, the Amadori compound in this case is lactulosyllysine and furosine is again a useful marker of lysine unavailability. For this reason several authors have used the furosine method for determining the progress of the Maillard reaction in different foods (Chiang, 1983; Hartkopf and Erbersdobler, 1993 and 1994, Henle et al, 1995; Resmini et al, 1990). However, today very powerful analytical techniques are disclosing new possibilities, permitting, for example, the direct determination of fructosyllysine (Vinale et al, 1999) by the use of a stable isotope dilution assay performed in liquid chromatography – mass spectrometry (LC–MS). This method overcomes the problems of hydrolytic instability of the analyte and the incompleteness of the enzymatic digestion technique. Other possible markers of lysine transformation are N-e-carboxymethyllysine (CML) and 5-hydroxymethylfurfural (HMF) (Fig. 11.10). CML was detected for the first time in milk by Büser and Erbersdobler (1986) and an oxidative mechanism was proposed for its formation (Ahmed et al, 1986). The formation of HMF in foods has been explained in two ways: via the Amadori products through enolisation (in the presence of amino groups) and via lactose isomerisation and degradation, known as the Lobry de Bruyn-Alberda van Ekenstein transformation (Ames, 1992). Because of this, it has recently been proposed to measure separately the HMF formed only by the acidic degradation of Amadori products and 274 The nutrition handbook for food processors CH2 O HO H H OH H OH CH2OH NH (CH2)4 H R'OC C NHR NH2 (CH2)4 H HOOC C NH2 O H2 C N H (CH2)4 O COOH NH2 H3C N (CH2)4 O NH2 COOH lysine furosine pyridosine Fig. 11.9 Compounds derived from fructosyllysine decomposition