Professional Documents
Culture Documents
Practica N°4 Extensión de Vida Útil en Productos de Panadería
Practica N°4 Extensión de Vida Útil en Productos de Panadería
• Revisar los principales mecanismos para extender la vida útil de productos panificados.
• Conocer el proceso industrial de fabricación de pan de molde.
• Relacionar los diferentes mecanismos para la extensión de la vida útil en términos de
conservación de las características texturales aplicados al proceso de fabricación de pan
de molde.
2. Procedimiento
III. CUESTIONARIO
1. Esquematice mediante un diagrama de flujo el proceso de producción industrial de
fabricación de pan de molde.
2. Describa los equipos y maquinarias que intervienen en cada etapa del proceso,
identificados en el diagrama de flujo.
3. Según los papers, como influye el proceso de retrogradación del almidón y sus
reacciones conexas en el endurecimiento del pan y en la velocidad con el que éste se
da.
4. Cuáles son los principales ingredientes que pueden ser utilizados para retardar el
proceso de endurecimiento del pan.
5. El pan de molde referido en el video tiene un tiempo de vida útil entre 30 – 35 días.
a. ¿Cuál es la razón para prolongar la vida útil de este pan por todo este tiempo?
b. Analizar la declaración de ingredientes en el empaque de cualquier pan de molde
de larga duración (Ejm: Bimbo, Pyc u otro) y con ayuda del Codex Alimentarius
identificar las sustancias y aditivos que fueron usados y clasificarlos por
funcionalidad.
c. Una vez identificados los ingredientes y aditivos responsables por la prolongación
de la vida útil haga una breve revisión bibliográfica de las funciones de cada uno de
ellos y en base a información de papers, especificaciones técnicas, hojas de
aplicación y/o recomendaciones de fabricante, trate de identificar las posibles dosis
para cada uno de ellos.
ABSTRACT: The molecular basis of staling is examined by reviewing what is known about the components of wheat flour,
factors that affect staling rate, and the various mechanisms that have been proposed. The conclusion reached is that bread
staling is a complex phenomenon in which multiple mechanisms operate. Polymer crystallizations with the formation of
supermolecular structures are certainly involved. The most plausible hypothesis is that retrogradation of amylopectin
occurs, and because water molecules are incorporated into the crystallites, the distribution of water is shifted from gluten
to starch/amylopectin, thereby changing the nature of the gluten network. The role of additives may be to change the
nature of starch protein molecules, to function as plasticizers, and/or to retard the redistribution of water between
components. Nothing more definite can be concluded at this time.
© 2003 Institute of Food Technologists Vol. 2, 2003—COMPREHENSIVE REVIEWS IN FOOD SCIENCE AND FOOD SAFETY 1
more readily retrograded fraction, amylose, was suspected first 2001). [Note: As pointed out above, the retrograded material
(see “Mechanisms of staling: Role of amylose” section). Evidence should have melted during baking so the effect would be one of
from Katz (1928) suggested formation of side-by-side associations adding corn amylopectin.]
of linear starch molecules in the B-type x-ray patterns of staled Toufeili and others (1999) found that an all-amylopectin Arabic
bread and retrograded starch. Hixon (1943) speculated that, if a bread (made with waxy barley starch and cross-linked waxy bar-
waxy wheat variety were available, then bread made from that ley starch in place of wheat starch) staled at a significantly faster
flour might not stale since it would be essentially void of amylose. rate than did Arabic bread made with normal wheat starch, that a
Alsberg (1927, 1928) pointed out the well-known fact that heat- low degree of starch crosslinking promotes recrystallization of
ing stale bread above 50 °C can restore the loaf to its original amylopectin [possibly by keeping polymer chains in close prox-
freshness. Since retrograded amylose will not melt at this tempera- imity to one another], and that a higher degree of crosslinking de-
ture (Knightly 1977), amylopectin was suggested to be the fraction creased the staling rate [possibly by restricting granule swelling
of starch responsible for staling. Supporting evidence was pre- and separation of polymer chains].
sented when bread prepared from a synthetic flour composed of Most agree that there is at least a correlation between amy-
waxy maize starch and nondevitalized gluten exhibited a normal lopectin retrogradation/crystallization and staling, even though
tendency to stale (Noznick and others 1946). Further, Schoch and the 2 events may not be part of the same process. Our conclusion
French (1947) found that the water-soluble material that could be is that amylopectin retrogradation is part of the staling process,
leached from bread crumb at 30 °C was predominantly amy- but is not solely responsible for the observed changes in texture.
lopectin. They hypothesized that progressive spontaneous aggre- For information on associations of starch polymer molecules in
gations of amylopectin molecules was responsible for bread firm- concentrated wheat starch (and other starch gels), see Keetels and
ing. Furthermore, they suspected that the contribution of the amy- others (1993, 1995, 1996 a,b,c,d; Vodovotz and others 2002).
lose fraction to staling was negligible, since they believed it to be Role of amylose. While Schoch and French (1947) believed
retrograded/insolubilized during cooling. The important role of that the linear fraction of starch had a negligible influence on
amylopectin in starch retrogradation was confirmed by calorime- bread staling, there is evidence that amylose is involved in some
try (Russell 1983a, b). way. Due to its rapid rate of retrogradation, Hoseney and others
However, Hoseney and Miller (1998) have pointed out that (1978) proposed that amylose was responsible for setting the ini-
stale bread must be heated to about 100 ºC before its compress- tial crumb structure, but not involved in the staling process. Er-
ibility approaches that of fresh bread (Ghiasi and others 1984) lander and Erlander (1969) theorized that amylose-amylopectin
and that, since retrograded amylopectin should have melted by aggregation was responsible for the changes that occur during ag-
the time the temperature reached 60 ºC, retrogradation of amy- ing of bread crumb. Kim and D’Appolonia (1977c) found that the
lopectin cannot be the only factor affecting firming. Retrograded solubility of amylose decreased markedly during the 1st d of
waxy corn starch (5%) was added to a bread formula and found bread storage, while the solubility of amylopectin decreased
to decrease gelatinization and to reduce the firming rate (Hibi steadily over 5 d of storage. They also found that the amount of
mechanism was not affected by protein content, suggesting that Biliaderis 1995; Biliaderis and Izydorczyk 1995; Cleemput and
the rate of staling is independent of protein quality. So they con- others 1997). Water-soluble and -insoluble pentosans have been
cluded that the primary effect of protein in reducing staling is dilu- reported both to retard staling and to have no effect on the staling
tion of starch. rate.
Several investigators have concluded that crumb firmness is not “Water-insoluble” pentosans. No differences in staling rate of
significantly correlated to flour protein type or concentration (Pon- breads made with or without tailings (starch fraction containing
te and others 1962; Leon and others 1997; Gerrard and others 9% water-insoluble pentosans) was observed by a sensory panel
2001). By examining a novel starch bread that contained no glu- (Bechtel and Meisner 1954b). Neither did Prentice and others
ten, Morgan and others (1997) suggested that starch retrograda- (1954) observe any effect on crumb firming rate due to tailings, al-
tion alone is sufficient to effect bread firming. though initial crumb firmness was decreased, probably due to the
Every and others (1998) suggested that, qualitatively, starch- high hydration capacity of pentosans. However, others found that
starch and starch-protein interactions are of equal importance to addition of water-insoluble pentosans resulted in a considerable
the staling mechanism, but that quantitatively, starch-starch inter- increase in loaf volume (Kulp 1968) and retardation of bread stal-
actions are more important since conventional wheat flour con- ing (Casier and others 1972, 1973; Denli and Ercan 2001). To
tains about 85% starch (db). They hypothesized that gluten is not add more confusion, addition of insoluble pentosans was report-
essential to the firming process and that increasing bread firmness ed to reduce bread quality, which could be overcome with addi-
results from chains of partially leached amylose and amylopectin tion of an optimum amount of pentosanase (Krishnarau and
attached to swollen, partially gelatinized starch granules interact- Hoseney 1994). Such variable results may result from differences
ing via hydrogen bonds with other starch granule remnants and, in type, molecular weight, and/or concentration of the pentosans
to a lesser degree, with gluten fibrils. Reconstitution experiments present in the formulation.
revealed that breads of equivalent specific loaf volume staled at “Water-soluble” pentosans. Contrary to the reports of less ben-
the same rate irrespective of protein type or concentration, but eficial effects of water-soluble pentosans (as compared to water-in-
other bread properties were altered by changes in the type or soluble pentosans), reports which were not confirmed by Kulp
concentration of protein (Gerrard and others 2001), lending sup- and Bechtel (1963) or Hoseney and others (1971), Michniewicz
port to the above hypothesis. and others (1992), like Jelaca and Hlynka (1972), found that wa-
Maleki and others (1980) postulated that the flour component ter-soluble pentosans had a significant positive effect on loaf vol-
primarily responsible for differences in staling rate is gluten and ume and that water-insoluble ones did not, that water-soluble
that its role in staling is something other than dilution of starch. pentosans retarded amylose aggregation, and that addition of wa-
Furthermore, they proposed that starch and water solubles were ter-insoluble pentosans decreased susceptibility of bread crumb
not involved significantly in determining the rate of staling. to a-amylase. They suggested that the contradictory results ob-
Martin and others (1991) proposed that bread firming is a result tained when studying the effects of pentosans on loaf volume may
of hydrogen bonding between gelatinized (partially pasted) starch have originated in differences in baking characteristics of the
granules and the gluten network in bread tying together the con- flours of various wheat cultivars, differences in chemical composi-
tinuous protein network and discontinuous granule remnants. tion of pentosans, and/or the way pentosans were incorporated
They theorized that the crosslinking interactions originate during into the dough. They further suggested that the reported reduction
baking; then during aging, the crumb loses kinetic energy, and in bread firmness upon storage when the dough was supplement-
both the number of interactions and their strength increases. ed with pentosans, as observed by Kim and D’Appolonia (1977d)
When reheated, bread freshness is restored because the and others, may have been a direct consequence of a higher
crosslinks (hydrogen bonds) and entanglements between gluten moisture content of the system.
and starch polymer molecules are easily broken. This theory is Interaction of pentosans with protein. It is possible that pen-
congruent with results of Dreese and others (1988), who reported tosans can interact with wheat-flour components other than
that starch and gluten molecules interact during baking. starch. Jelaca and Hlynka (1972) proposed that pentosan-gluten
Gerrard and others (1997) suggested a modification to the hy- interactions were responsible for baking improvement effected by
pothesis of Martin and others (1991). They agreed with the hy- pentosans. Based on the effects of actions of arabinoxylanases,
pothesis that staling is a result of increasing interactions between Cleemput and others (1997) suggested that there are associations
swollen starch granules and the gluten network. However, they of arabinoxylans with proteins and/or other wheat components in
put forth the opinion that the decrease in firming rate in breads doughs.
made with a-amylase (see “Enzymes: ␣-amylases and debranch- Pentosans and starch retrogradation. The effect of pentosans
ing enzymes” section) as a dough additive is not the direct result on starch retrogradation has been investigated using both starch
of starch hydrolysis products (dextrins and maltooligosaccha- gels and bread itself. Gilles and others (1961) reported that water-
rides), some of which, they suggest, are nonspecifically associated soluble pentosans found in the “soluble starch” extract of bread
with the protein matrix, but a result of modification of swollen crumb inhibited retrogradation of amylose and that, although the
starch granules in such a way that their interaction with the pro- pentosans affected some characteristics of the bread, staling rate
tein network is reduced (presumably either qualitatively or quanti- was not one of those characteristics. Kim and D’Appolonia
tatively). (1977a) found that pentosans had a definite effect on retarding
Rogers and others (1988) reported that, even though shortening starch retrogradation in wheat starch gels, with the effect of water-
and native lipids have significant effects on bread staling, neither insoluble pentosans being more pronounced. They reported that
have major effects on starch retrogradation. They suggested for- water-soluble pentosans reduced retrogradation by acting on
mation of protein-lipid interactions. amylopectin, while water-insoluble pentosans reduced the degree
Role of pentosans. As mentioned in the “Nonstarch polyosac- of retrogradation of both amylose and amylopectin. Similar results
charides” section, the influence of the so-called pentosans on were found when the effect of pentosans on staling was studied in
breadmaking and bread properties is not clear, although the sub- a bread system (Kim and D’Appolonia 1977d). Results indicated
ject has been examined extensively (Kulp 1968; D’Appolonia that the basic mechanism of bread staling was unchanged; thus, it
1971, 1980; Hoseney 1984; Meuser and Sukow 1986; Jank- was suggested that pentosans decreased the staling rate by reduc-
iewicz and Michniewicz 1987; Roels and others 1993: Rattan ing the amounts of starch components available for retrogradation
and others 1994; Krishnarau and Hoseney 1994; Izydorczyk and (Kim and D’Appolonia 1977d). However, others have concluded
Vol. 2, 2003—COMPREHENSIVE REVIEWS IN FOOD SCIENCE AND FOOD SAFETY 5
amylopectin crystallization”. leased from gluten into crystalline structure of starch that devel-
Crumb-crust redistribution of moisture. As baked bread be- oped upon staling. [Note: The B structure has 36 water molecules
gins to cool, a moisture gradient forms in the loaf (Piazza and in the unit cell, whereas the A structure has only 8 (Sarko and Wu
Masi 1995). Differences in vapor pressures between the crust and 1978).] Conversely, Kim-Shin and others (1991) proposed that the
the internal region of the loaf result in moisture migration from the redistribution of water occurs in the amorphous phase. The ratio
crumb to the crust (Stear 1990). Over time, the moisture content of starch to gluten (6:1) in bread crumb ensures that moisture
in the center of the loaf decreases, while that in the external region transfer to the starch would result in firming of the continuous glu-
increases (Bechtel and others 1953). ten phase (Willhoft 1971). It is important to keep in mind at all
Baik and Chinachoti (2000) found that bread stored with its times, however, that the change in the state of water cannot be
crust became significantly firmer than bread stored without its correlated directly to the retrogradation process (Wynne-Jones
crust and contained more recrystallized amylopectin, indicating and Blanshard 1986).
that moisture redistribution from crumb to crust plays a significant Levine and Slade (1990) and Slade and Levine (1991) present
role in firming, a conclusion confirmed by a loss in freezable wa- thorough and well-documented evidence for the role of water in
ter in the crumb of bread stored with crust, which correlated with the staling process. Their arguments are based upon the mecha-
changes in its thermomechanical profile. nism of polymer crystallization, polymer crystallization kinetics as
Several NMR parameters correlate with crumb firming and are a function of glass transition and melting temperatures, water as a
believed to be related to both microscopic and macroscopic re- plasticizer, and sugars as antiplasticizers in the system. In their re-
distribution of water (Chen and others 1997b). Using NMR tech- view, Slade and Levine (1991) state essentially that “if adequate
niques, it has been found that, as staling proceeds, the water in packaging prevents simple moisture loss, the predominate mecha-
bread becomes less and less mobile (Leung and others 1983; nism of staling in bread crumb is the time-dependent recrystalliza-
Wynne-Jones and Blanshard 1986; Kim-Shin and others 1991; tion of amylopectin from the completely amorphous state of a
Chen and others 1997a,b; Engelsen and others 2001). However, freshly heated product to the partially crystalline state of a stale
Ruan and others (1996), using MRI, found that, as storage time of product, with concomitant formation of network junction zones,
sweet rolls increased, mobility of the less-mobile water fraction redistribution of moisture via both microscopic and macroscopic
decreased, while mobility of the more-mobile fraction increased. migration (Czuchajowska and Pomeranz 1989), and increased
Moisture redistribution among components. Transfer of mois- textural firmness (Kulp and Ponte 1981; Russell 1983b; Russell
ture from one constituent of the bread crumb to another is general- 1987).” They further point out that there is evidence from studies
ly accepted as a contributing factor in staling, possibly being re- of starch gels/pastes that the rate and extent of amylopectin crys-
sponsible for the perceived dryness of stale bread (Senti and Dimler tallization depends on the mobility of its outer branches (Ring and
1960). Water is a plasticizer, making the bread components more others 1987; Russell 1987; Marsh and Blanshard 1988; Slade
flexible. Thus, as water is removed (from either gluten or starch or and Levine 1989, 1991) and on sample history, since the pro-
both), increasing crumb firmness should occur. Whether staling in- cesses that occur both during heating/baking and during aging/
volves dehydration of gluten or starch has been studied extensively, storage are nonequilibrium processes (Ring and others 1987;
but is still unclear. However, the majority of evidence suggests a Slade and Levine 1989, 1991). [Note: Slade and Levine refer to
gluten to starch transfer of water as the starch crystallizes. recrystallization of amylopectin, and indeed it is a recrystalliza-
Katz (1928) first suggested that, during staling, moisture was re- tion. We have not used the term elsewhere in this review so as to
leased from starch and taken up by gluten. Senti and Dimler (1960), make it clear that amylopectin molecules do not recrystallize to
by studying equilibrium relative humidities, also suggested that the same crystalline state that they were originally in nongelati-
moisture transfer would likely occur from starch to gluten. Cluskey nized granules.]
and others (1959) reported a progressive drop in moisture-sorption Amylopectin crystallization results in a partially crystalline, su-
capacity for starch and lack of a change for gluten, indicating a permolecular structure containing disperse B-type crystalline re-
transfer of moisture from starch to gluten during aging. gions (Slade and Levine 1987). Incorporation of water molecules
In contrast, Alsberg and Griffing (1927) and Alsberg (1936) pos- into the crystal lattice occurs during formation of the B-type poly-
tulated that it was the gluten that hardened as result of moisture morph (Imberty and Perez 1988) and, thus, a redistribution of
loss to starch. This concept is supported by data of Bachrach and moisture is effected. This process was demonstrated by a progres-
Briggs (1947), who observed an increase in moisture-sorption ca- sive decrease in the percentage of “freezable” water as bread was
pacity of gelatinized starch upon aging [contrary to the results of stored over 11 d (Slade and Levine 1991). The water molecules
Katz (1928) and Cluskey and others (1959)]. Further evidence that are part of the crystal lattice are not available for plasticiza-
came from investigations by Willhoft and coworkers (Breaden and tion, so the result is the perceived drier, firmer texture characteris-
Willhoft 1971; Willhoft 1971; Kay and Willhoft 1972), who re- tic of stale bread. So, all in all, amylopectin crystallization in bread
ported that gluten undergoes a 1st-order transformation resulting requires both microscopic and macroscopic redistribution of wa-
in the release of water from gluten and absorption of this water by ter so that there is sufficient moisture present at the locus where
retrograding starch. crystallization takes place to plasticize polymer chains so that
The notions of “free” and “bound” water have been reported to they are mobile enough for crystallization to occur and for incor-
be of importance in altering the rate or extent of staling in bread poration into B-type crystal latices (Levine and Slade 1990; Slade
(Knjaginciev 1970). More recently, the use of NMR and a greater and Levine 1989, 1991).
understanding of the role and mechanism of starch polymer crys- It seems clear that moisture transfer between bread compo-
tallization have led to the conclusion that starch takes up water nents, specifically between gluten and starch, occurs as bread
from gluten upon aging of bread. Leung (1981) and Leung and ages. However, like other measurable changes in the nature of
others (1983) proposed that, as starch changes to a more crystal- bread components, the role, if any, of moisture and moisture re-
line state, more water molecules become immobilized due to their distribution in the staling process remains undetermined. (See
incorporation into crystal structures. Chen and others (1997a,b) also “Mechanisms of staling: Role of pentosans” and “Carbohy-
reported a decrease in water mobility in bread upon staling, in drate ingredients” sections).
agreement with results of others (Wynne-Jones and Blanshard
1986; Slade and Levine 1991), and concluded that the decrease Processing factors
in water mobility was due to incorporation of water molecules re- Effects of technological factors, which include manufacturing
Vol. 2, 2003—COMPREHENSIVE REVIEWS IN FOOD SCIENCE AND FOOD SAFETY 7
duced mainly low-molecular-weight, branched dextrins of DP 19- their presence is simply correlated with a key modification of
24 that either had less ability to retrograde, interfered with amy- starch granules that is related to reduced staling, possibly by re-
lopectin retrogradation, or interfered with whatever other interac- ducing gluten-starch interactions. Duedahl-Olesen and others
tions are responsible for crumb firming. (1999) also reported that maltooligosaccharides of average DP up
Duran and others (2001) attributed the antistaling effect of ␣- to 20 had no effect on formation of the staling endotherm. Inter-
amylases to the production of maltooligosaccharides. Biliaderis estingly, they did find that amylopectin recrystallization was re-
and Prokopowich (1994) found that maltose and maltotriose had duced significantly when ␥-cyclodextrin (3%) was incorporated.
antiretrogradation effects on starch gels and proposed that the Slade and Levine (1987) and Levine and Slade (1990) studied
chain ordering of amylopectin in sugar-containing starch gels is a maltooligosaccharides of DP 3 to 8 at relatively high concentra-
function of the compatibility of the sugar with the structure of wa- tions (1:1, oligosaccharide:water). Their conclusion was that the
ter. Solutes that fit well in the water structure retarded chain reor- reported antistaling effect could be explained by an impact on the
dering. On the other hand, solutes that disturb water structure Tg, resulting in a smaller ⌬T above Tg, which retarded the starch
promoted ordering and aggregation of starch molecules. Maltotri- crystallization process. Further, they reported a relationship be-
ose, which was reported to be the most effective maltooligosac- tween an increase in Tg and the degree of staling (Slade and Le-
charide in impeding retrogradation, disturbs the structure of water vine 1991). The correlation of an increase in Tg during staling with
only slightly (Biliaderis and Prokopowich 1994). the firming of bread was confirmed by Jagannath and others
Defloor and Delcour (1999) reported that starch hydrolysis (1999a).
product preparations with average DPs of from 4 to 66 reduced Dragsdorf and Varriano-Marston (1980) studied the effects of
DSC staling endotherms in baked and stored bread doughs. They barley malt, fungal ␣-amylase, and bacterial a-amylase on starch
attributed their antistaling effect to a reduction in starch recrystalli- crystallization and organization in staling breads using x-ray dif-
zation but did not speculate about a mechanism. fraction (see also Akers and Hoseney 1994). Comparing stored
Martin and Hoseney (1991) proposed that low-molecular- and fresh breads, they found that the degree of crystallinity of
weight dextrins (maltooligosaccharides) produced by a-amylases breads baked with different enzyme sources was in the order bac-
were directly responsible for the antistaling phenomenon ob- terial ␣-amylase > cereal ␣-amylase > fungal ␣-amylase > control.
served by enzyme addition. Their explanation was that the low- These results were in opposition to bread firming data, suggesting
molecular-weight products inhibited cross-link formation between to them that starch crystallization and bread firming are different
starch and gluten. and separate processes. Their results agreed with those obtained
Min and others (1998) studied the effect of 2 novel antistaling by Zobel and Senti (1959), who suggested that bacterial ␣-amylas-
amylases. When added to bread, they produced selectively either es inhibit staling by breaking interconnecting chain associations
maltose and maltotriose or maltotetraose and maltotriose. Based in the network of starch crystallites.
on the results, they postulated that maltotriose and maltotetraose Retardation of bread staling, while avoiding a gummy mouth-
were directly responsible for retarding retrogradation in bread, feel, was achieved by incorporating pullulanase with a cereal or
suggesting that these oligomers were of the right size to interfere bacterial ␣-amylase in the dough (Carroll and others 1987). A
with starch-gluten interactions [theory of Martin and others (1991) product produced by action of pullulanase or isoamylase on
and Martin and Hoseney (1991) on the mechanism of staling]. starch, which the inventors refer to as low-molecular-weight amy-
Maltose was found to be less effective in bread staling prevention, lose, but which is in reality a mixture of released branch chains,
and it was suggested that its relatively small size and its ability to when added to a dough formulation, was reported to have an an-
diffuse easily might be the reason why it was less effective than tistaling action (Yoshida and others 1972).
maltotriose or maltotetraose (Min and others 1998). Donnelly and All in all, it appears that starch hydrolysis products are involved
others (1973) reported that there is a slight decrease in moisture in inhibition of staling, but that the products must be of a unique
adsorptive capacity as the molecular size of maltooligosaccha- type, perhaps either maltotriose and maltotetraose or products
rides increases from DP 3 to DP 11, and that maltose was the ex- larger than those present in traditional maltodextrin preparations.
ception, being less hygroscopic than was the DP 11 maltooli- However, that the presence of such products is only correlated to
gosaccharide. This led Min and others (1998) to conjecture that some other modification in starch (or another component) that is
maltotriose and maltotetraose might hold water around starch the real determinant cannot be ruled out.
molecules and inhibit starch-starch interactions more than mal- Lipases. Although Johnson and Welch (1968) patented lipase
tose does. formulations that retard staling in bread, the use of lipases for
Despite conclusions that dextrins directly affect staling in bread, breadmaking was virtually unknown until recently (Qi Si 1997).
considerable evidence has been published to the contrary. Salem Depending on the type of flour and the formula, addition of some
and Johnson (1965) found, from experiments in which starch hy- 1,3-specific lipases resulted in more uniform crumb structure and
drolysis products were added to a bread dough formula, that cer- thus an improvement in crumb softness during storage (among
tain maltooligosaccharides (such as maltohexaose and -heptaose, other dough conditioning improvements). Furthermore, these li-
as compared to glucose, maltose, and maltotriose, -tetraose, and - pases were shown to be a replacement for shortening, although
pentaose) and dextrins increased the rate of crumb firming, in no improvement in crumb elasticity was found.
contrast to results obtained when ␣-amylase was incorporated as Siswoyo and others (1999) found that, while use of a purified li-
an additive. However, in contrast, Every and others (1992) found pase alone retarded retrogradation in bread crumb, use of a com-
that maltooligosaccharides of DP 3-10 correlated with a reduction bination of a purified lipase and a purified a-amylase reduced ret-
in firming rate, and Akers and Hoseney (1994) implied that starch rogradation to a much greater extent. [Note: Since commercial en-
hydrolysis products of a size greater than maltoheptaose might zyme preparations are rather crude and probably contain both
have antifirming properties. activities, this combination could unknowingly be involved in the
There is a 3rd conclusion. Because added maltooligosaccha- antistaling activity.]
rides did not survive fermentation and because the presence of Qi Si (1997) suggested that the mechanism of retrogradation re-
maltooligosaccharides of a specific size class could not be corre- tardation does not involve the most obvious explanation: hydrol-
lated with the firming rate of bread, Gerrard and others (1997) ysis of lipids to monoglycerides, which are reported to have anti-
concluded that maltooligosaccharides (DP 3-8) produced by ␣- staling characteristics. [Note: Tri- and diacylglycerols do not de-
amylases are not themselves responsible for antistaling, but that crease crumb firmness, but monoacylglycerols/monoglycerides
Vol. 2, 2003—COMPREHENSIVE REVIEWS IN FOOD SCIENCE AND FOOD SAFETY 9
dition of monoglycerides may counteract staling of breads during vestigations by Ponte and Titcomb (1971), Kulp and Ponte (1981)
storage, an increased tendency to crumble may result (Malkki and concluded that a surfactant’s ability to retard firming is more im-
others 1978). portant than an initial softening of crumb in freshly baked bread.
The mechanism of the antistaling effect of monoglycerides is Amylose complexes. Details of fatty acid-amylose complexes
still unknown, but it is thought to be different from that of shorten- have been examined using x-ray diffraction, DSC, and electron
ing (Rogers and others 1988), for monoacylglycerols can replace microscopy (Godet and others 1993b, 1995a, 1996). Using mo-
shortening, but shortening cannot replace monoacylglycerols. lecular modeling techniques, Godet and others (1993a,b, 1995b)
Krog and others (1989) concluded that reductions in crumb concluded that the hydrocarbon tails of complexed fatty acid
firmness brought about by addition of monoglycerides were molecules are indeed inside the hydrated V helix (see below) with
probably the result of interactions with amylose rather than with the polar head group outside the lumen. Interactions of amylose
amylopectin. When relatively large amounts of monoglycerides with over 20 surfactants were studied, and a “complexing index”
are used, essentially all released amylose can be complexed (as was calculated and assigned to each one (Krog 1971). Morad and
measured by DSC); interactions with amylopectin are also in- D’Appolonia (1980) demonstrated that incorporation (0.5%) of 5
creased. At lower concentrations, monoglycerides interact prima- commercial surfactants resulted in amylose-surfactant complexes.
rily with amylose because of competition between the 2 poly- It was found by Eliasson (1985) that the amount of amylose
mers. leached from starch granules decreased in the presence of emul-
Polyoxyethylene monostearate (POEMS). POEMS, a reaction sifiers.
product of ethylene oxide and stearic acid, was one of the first ad- Numerous studies have dealt with the ability of polar lipids to
ditives reported to retard staling (Maga 1975). Favor and Johnson inhibit bread staling. Mikus and others (1946) suggested that a he-
(1947) demonstrated that POEMS (0.5 to 1.0%) dramatically re- lical complex formed between amylose and MG, thus effecting a
duced the firming rate of bread between the 1st and 3rd d. Other softer crumb, but without affecting the firming rate. Schoch (1965)
results (Freilich 1948; Edelmann and Cathcart 1949; Edelmann reached a similar conclusion.
and others 1950; Skovholt and Dowdle 1950) confirmed that PO- Data from Lagendijk and Pennings (1970) provided evidence of
EMS was effective in reducing the rate of firming. Carson and oth- the relationship between amylose-lipid complexation and the in-
ers (1950) theorized that POEMS retarded staling by 2 mecha- hibition of staling. They reported maximum complexation with
nisms: (1) by insolubilizing amylose and (2) by interacting with monopalmitin, which corresponded with the softest crumb after
starch granules via hydrogen bonding. 48 and 72 h of storage and concluded that complexation reduces
Sodium stearoyl lactylate (SSL). Pisesookbunterng and the flexibility of amylose molecules and thereby reduces their ret-
D’Appolonia (1983) found that, among various surfactants stud- rogradation.
ied, SSL had the greatest binding affinity to starch. The anionic sur- Pisesookbunterng and D’Appolonia (1983) reported that sur-
factant might also prevent protein denaturation. Calcium stearoyl factants (SSL, MDG, and 40% Poly-60 / 60% MDG blend) ad-
lactylate is less effective as a crumb softener, but is active. sorbed to the starch granule surface, preventing moisture uptake
Glycerol monostearate (GMS). GMS is used in many starch- by the starch from gluten during aging of bread. However, water
based food products to improve physical characteristics, includ- was able to migrate from crumb to crust. Firmness of fresh bread
ing the degree of softness after storage (Krog 1971). was not affected by the surfactant, although firming rate during
Other surfactants. Other surfactants that are effective as anti- storage was slowed. Xu and others (1992) confirmed these results.
staling agents include polyoxyethylene sorbitan monostearate On the other hand, no apparent relationship between amylose-
(Polysorbate 60), succinylated monoglycerides, and glycerol. surfactant complex formation and reduction of crumb firming was
Novel surfactants or surfactant blends have been formulated for found by Osman and others (1961).
use as antistaling agents in bakery products. A blend developed X-ray diffraction can be used to detect complex formation, as
by Knightly (1987) consisted of a hydrophilic lecithin and at least well as crystallinity and crystal types in general (Zobel 1973). The
one of the following: monoglyceride, lactic acid esterified amylose V complex helix hydrate can be detected by measuring
monoglyceride, succinic acid esterified monoglyceride, maleic the intensity of the characteristic 4.4Å diffraction line. Formation
acid esterified monoglyceride, or edible salts of stearoyl lactylic of the B-type crystal structure is followed by measuring the inten-
acid. This blend was claimed to both inhibit staling and to act as a sity of the 5.25Å spacing. The B structure, typical of retrograded
dough conditioner. Other antistaling surfactant blends were de- starch, is extended, unlike the tight V-form helix. Molecules in
veloped by Vidal and Gerrity (1979). these 2 forms do not cocrystallize (Zobel and Senti 1959). With
Mechanism of antistaling effect of surfactants. The mechanism no surfactants present, bread that is freshly baked shows only V-
by which surfactants influence crumb firmness has been debated crystallinity due to amylose-lipid complexes formed with the na-
and is discussed briefly in the following sections. Amylose-surfac- tive fatty acids in the starch granules (Zobel 1973), during dough
tant, amylopectin-surfactant, and protein-surfactant interactions heating (Zobel and others 1988). Thus, a portion of the amylose is
have all been investigated, as has starch swelling in the presence insolubilized during the overall baking process. The remaining
of added surfactants. Whether surfactants actually decrease the pattern is an amorphous halo produced by gelatinized starch.
rate of firming or produce softer breads that then stale at the same During bread aging, the amorphous starch crystallizes into (B-
rate as the control has been debated. Surfactants have multiple type) crystals, while the V-type intensity (amylose-lipid complexes)
properties, resulting in multiple functionalities, so definitive exper- remains unchanged (Zobel and Senti 1959). With surfactant add-
iments examining a cause-and-effect relationship with regards to ed, breads were softer after 3 d as measured by a compressimeter
staling are difficult, if not impossible, to design. (Zobel and Senti 1959). V-lines increased with surfactant addition,
In excess water, surfactants do not change the gelatinization indicating complexation between amylose and the adjuncts. But,
temperature, but they do delay pasting (Miller and others 1953). once again, complexation does not necessarily correlate with sur-
Whether this is related to their functionality in breadmaking is un- factant effectiveness in retarding staling (Osman and others 1961;
known. Knightly (1977) reported that surfactants had little to no Zobel 1973).
effect on initial crumb firmness, but did affect the firming rate dur- Dragsdorf and Varriano-Marston (1980) suggested that surfac-
ing storage, a finding in agreement with earlier reports (Favor and tants that produce a V complex hydrate structure (for example,
Johnson 1947; Skovholt and Dowdle 1950; Hopper 1949; Edel- SSL) may either prevent migration of starch polymer molecules
mann and others 1950). Based on unpublished results from in- from granules during baking, thus retarding crumb firming, or re-
Vol. 2, 2003—COMPREHENSIVE REVIEWS IN FOOD SCIENCE AND FOOD SAFETY 11
ty. They also found that all the hydrocolloids they used generally effectively than did monostearin (Watanabe and others 1981).
improved quality and that those with higher water-holding capaci- Propylene glycol was found to be superior to glycerol, malto-
ty increased crumb firmness. In contrast, Davidou and others dextrins, gelatin, commercial ␣-amylase, and poly(propylene gly-
(1996) reported that both degrees of crumb firmness and the rate col) in antistaling activity (Jagannath and others 1998). 1,3-Bu-
of staling during storage were reduced by addition of locust bean tanediol and 1,3-heptanediol (0.5%) reduced the staling rate of
gum, alginate [presumably sodium alginate], and xanthan. They bread significantly, with 1,3-butanediol having the greatest effect
proposed that the gums modified the organization of the amor- (Frankenfeld and others 1977).
phous part of the crumb, perhaps by inhibiting gluten-starch inter- Incorporation of durum wheat flour (25%) into a bread wheat
actions, perhaps in the same manner as proposed for dextrins flour did not improve initial firmness, but did retard staling
(Martin and others 1991). They also reported that only locust through 4 d of storage (Boyacioglu and D’Appolonia 1994).
bean gum (of the 3 gums) effected water retention. Carboxymeth- Breads made with triticale flour staled twice as fast as did bread
ylcellulose (CMC) and hydroxpropylmethylcellulose (HPMC) made with wheat flour (Tsen and others 1973).
(0.3%) also decreased initial firmness (Armero and Collar 1998). Sato and others (1989) reported staling retardation during long-
No increases in the Avrami value were found. Hydrocolloid-glu- term storage when methyl 3,6-anhydro-␣-D-glucopyranoside was
ten entanglements or linkages were suggested. added to the formula.
Addition of psyllium husk powder/gum (2, 4, or 8%) decreased
the staling rate of bread as measured by compressibility and DSC
(Czuchajowska and others 1992). Moisture content remained Flavor Changes
constant. Bread softness improved without increasing the possi- Staled bread is considered unacceptable due to changed flavor.
bility of microbial deterioration. Lent and Grant (2001) found that A review on bread flavor is available (Lorenz and Maga 1972).
bagels containing added xanthan had slightly higher crumb mois-
ture contents and staled at a somewhat reduced rate (DSC). Addi-
tion of pectin increased the specific volume of bread and reduced Processes for Acceleration of Staling
the rate of firming during storage (Kegoya-Yoshino 1997). A process by which the staling rates of bread products can be
Replacement of 10% of the wheat flour with steamed oat flours accelerated using a time-temperature-moisture protocol to pro-
retarded bread staling without adversely affecting the loaf volume duce croutons and dry crumbs at a faster rate has been patented
(Zhang and others 1998). The reduction in the rate of staling was (Slade and others 1987). The staling process is accelerated via
attributed to the high water absorption capacity of the -glucan in temperature cycling. Bread is exposed alternately to a temperature
oat flour, but oat starch has been found to retrograde at a slower just above the glass transition temperature (maximum rate of nu-
rate compared to other starches (White and others 1989). cleation) and to a temperature just below the melting temperature
Patents have been issued for the use of karaya gum (Andt 1966) (maximum rate of crystal growth). Other patents for preparing
and what is called low-molecular-weight amyloses, but which in bread crumbs and/or croutons have been developed (Tu and oth-
reality are the branch chains of amylopectin released by the ac- ers 1986; Dyson and others 1980).
tion of an a-1,6-glucan hydrolase as antistaling agents (Yoshida
and others 1972).
It has also been reported (although not supported in the article Summary of the Basis of Staling and Factors Affecting the
with published experimental data) that methylcellulose and hy- Rate of Staling
droxpropylmethylcellulose extend the shelf life of baked products Bread staling is a complex phenomenon, certainly involving
via prevention of water loss during baking (Dziezak 1991). The multiple factors. Much has been learned about bread staling, and
same report states that guar gum and xanthan gum function as an- application of this knowledge has led to considerable improve-
tistaling agents. ments in shelf life. However, without knowledge of the precise
Damaged and modified starch. Modified starches have been mechanism, addressing the problem of bread staling remains a
investigated as antistaling agents (Maga 1975). Tipples (1969) re- process of formulating and testing more and more hypotheses. It
ported that the use of 25 to 35% damaged wheat starch de- is difficult to determine cause-and-effect relationships because in-
creased the rate of staling, especially when malt was added and volvement of a constituent may be indirect and additives, other
the sponge-dough method was used. changes in formulation, and process changes may alter more than
On the 4th d of storage, breads containing 5% of a phosphory- one property and the effects may cancel each other.
lated waxy maize starch were as fresh as a 1-d-old control bread Retrogradation of starch molecules remains the most widely ac-
(Bergthaller and Stephan 1970). The water-holding capacity of the cepted factor contributing to bread staling, but it must be remem-
bread was not affected by the starch phosphates. bered that there is also good evidence that there is no cause-and-
effect relationship between retrogradation and staling. While amy-
Miscellaneous lopectin retrogradation is believed to play the major role, amylose
Results of use of dairy ingredients in breads for antistaling pur- is now also thought to be involved. And while amylose-surfactant
poses have been inconsistent (Mannie and Asp 1999). complex formation has been a widely used strategy for reducing
D’Appolonia (1984) reported that milk solids have little to no ef- bread staling, amylopectin complexes may also be important, not
fect on bread staling, but do soften the crumb initially. Conversely, necessarily related to an inhibition of retrogradation because ad-
others have suggested that nonfat dry milk solids retard staling ditives that retard starch retrogradation may not retard staling.
(Dubois and Dresse 1984). Acidic whey (concentrated or uncon- Moisture content and moisture transfer among bread compo-
centrated) retarded staling in Hamam (French-type) bread at 1% nents is believed by many to be a significant factor contributing to
whey solids (Yousif and others 1998). Neither acid casein or sweet bread staling. Most evidence supports the concept that gluten
whey powder were found to reduce staling in bread significantly serves as a moisture reservoir from which water is transferred to
(Erdogdu-Arnoczky and others 1996), while acid whey powder retrograding starch molecules. But the relative effects of dehydra-
did. Despite its high water-holding capacity, succinylated whey tion of gluten and hydration of starch can only be surmised, as
protein concentrate did not prevent bread staling (Thompson and can the degree of benefit from prevention of this type of moisture
Baker 1983). transfer.
L-Leucine n-alkyl esters slowed the staling rate of bread more Evidence has accumulated for a major role of gluten in bread
Vol. 2, 2003—COMPREHENSIVE REVIEWS IN FOOD SCIENCE AND FOOD SAFETY 13
Amylo-Graph, Rapid Visco Analyzer, and related instruments Isothermal microcalorimety, a technique which is much more
have been used to measure the extent of starch gelatinization in sensitive and requires larger samples sizes than does convention-
bread crumb slurries (Yasunaga and others 1968). Peak viscosity al DSC (Karim and others 2000), has been used to study the early
changes were suggested as an index of staling because it was stages of starch retrogradation and has been demonstrated to be
thought that peak viscosity would decrease with age due to a effective for examination of the antistaling effects of lipids and sur-
toughening effect on partially gelatinized starch granules during factants (Silverio and others 1996).
staling. Based on results in which the outer 1-cm and 2-cm por- Other thermoanalytical instruments include thermogravimetric
tions of the crumb produced a lower peak viscosity than did the analysis (TGA), thermomechanical analysis (TMA), and dynamic
center portion, it was concluded that starch granules in the crumb mechanical analysis (DMA). TGA measures changes in the weight
center were less gelatinized than those in the crumb exterior. De- of a sample as a function of temperature (Schenz and Davis
spite reporting amylograph data that agreed with those of Yasuna- 1998). While events such as volatization, dehydration, and chem-
ga and others (1968), Varriano-Marston and others (1980) con- ical reactions can be observed using TGA, other simple transitions
cluded that the amylograph does not indicate the degree of starch can be missed if no weight changes occur (Sperling 1992).
swelling accurately in bakery products, but rather shows the sum Schiraldi and others (1996b), using TGA, found that the release of
of the contributions of all macromolecules to the viscosity of the water upon heating bread corresponded to 2 main binding states,
bread slurry. Toufeili and others (1994) found that, as staling in- and that the 2 fractions were dependent on the age of the bread.
creased, pastes made from Arabic bread changed from being vis- TMA measures changes in penetration, extension, expansion,
coelastic solids (G” < G’) to elastoviscous liquids (G” > G’). or contraction as a function of temperature (Schenz and Davis
1998) and can be used to determine the Tg of a substance by de-
Thermal analysis tecting a change in the thermal expansion coefficient (LeMeste
Thermal analysis has been used extensively to study starch ret- and others 1992). The deformation of a substance is measured
rogradation as well as bread staling (Russell 1983a, b; under nonoscillatory (static) load as the substance is subjected to
Czuchajowska and Pomeranz 1989; Le Meste and others 1992; a controlled temperature program (Flynn 1990). LeMeste and oth-
Schiraldi and others 1966a,b; Champenois and others 1995; ers (1992) developed a TMA method to measure the glass transi-
Vodovotz and others 1996; Baik and Chinachoti 2000), and its tion of white pan bread.
use has been mentioned throughout this review. Of the thermoan- DMA measures the dynamic moduli and damping of a sub-
alytical methods, differential scanning calorimetry (DSC) and dif- stance under oscillatory load as a function of temperature and fre-
ferential thermal analysis (DTA) have proven to be the most useful quency as it is subjected to a controlled temperature program
in providing basic information on starch retrogradation (Karim (Flynn 1990). DMA has also been referred to as forced oscillatory
and others 2000). Because both measure the differential tempera- measurements, dynamic mechanical thermal analysis (DMTA),
ture or heat flow to or from a sample versus a reference material dynamic thermomechanical analysis, and dynamic rheology (Me-
as a function of time, both can be used to monitor such changes nard 1999). In DMA, as oscillatory stress is applied to the sample
as phase transitions, molecular conformational changes, interac- in the bending or tensile mode of deformation, the lag of the re-
tions with other components, and pyrolytic degradation of the sulting oscillatory strain is measured. DMA is 1000 times more
sample. Specialized DSC instruments, including modulated DSC sensitive in observing thermal transitions than is DSC (Vodovotz
and polarization DSC, are also available (Schenz and Davis and others 1996). DMA has been used to study staling profiles of
1998). Indian unleavened breads by Jagannath and others (1999b), to in-
When aged bread samples are heated in a DSC pan, an endot- vestigate the effects of added hydrocolloids, pentosans, and solu-
herm is observed as reordered amylopectin reaches its glass tran- ble proteins on bread staling (Schiraldi and others 1996a), and to
sition and/or melting temperature, and the enthalpy change asso- examine the effect of aging and drying on thermal transitions of
ciated with this transition can be measured. Because the time bread (Vodovotz and others 1996).
scales for endotherm development and for the increase in crumb
firmness are broadly similar in magnitude, DSC can be used to Infrared spectroscopy
measure the rate of bread staling quantitatively (Jagannath and Fourier transform infrared (FTIR) spectroscopy and near-infrared
others 1999a). However, there are overlapping transitions over a (NIR) spectroscopy, which have the advantage of being noninva-
wide temperature range because of the variety of components sive methods, have been used to monitor staling in bread (Wilson
and range of structures present, which cause difficulty in analysis and others 1991).
(Vodovotz and others 2001). Fourier transform infrared (FTIR) spectroscopy. Because FTIR
DTA was used to investigate bread staling by Axford and Col- spectroscopy measures the degree of short-range ordering in a
well (1967). An endotherm peak, which was absent in fresh bread system, conformational changes brought about by starch retrogra-
samples, developed during storage, and the increases in peak dation can be monitored by analyzing the band-narrowing,
area were proportional to increases in bread firmness (Cornford which is caused by a reduction in the range of conformations and
and others 1964). Because an increase in glass transition temper- a smaller distribution of bond energies due to the system becom-
ature (Tg) of bread crumb stored for different times was correlated ing more ordered upon staling (Wilson and others 1991; Karim
(96.53%) with an increase in the degree of bread staling as mea- and others 2000). Changes in band intensities in the 1300 to 800
sured by compression analysis, it was concluded that the mea- cm-1 region correlate to conformational changes during starch ret-
surement of Tg during storage could be used to quantitatively pre- rogradation. Peaks at 1047 cm–1, which relate to crystalline re-
dict the rate of staling (Jagannath and others 1999a). DSC studies gions of starch, and at 1022 cm–1, which are characteristic of
of starch can approximate gelatinization during baking, since in amorphous regions of starch, are of particular interest (Karim and
both cases the gelatinized starch granules are swollen, but non- others 2000). Thus, starch retrogradation can be defined (in terms
disrupted (Jacobson and BeMiller 1998). Thus, the conditions of of FTIR data) as an increase in the ratio of peak intensities at 1047
gelatinization in the calorimeter more closely approximate those and 1022 cm–1 (Smits and others 1998).
encountered during baking than those encountered during starch Near infrared (NIR) reflectance spectroscopy. Radiation scat-
pasting. Unlike compressibility measurements, endotherm peak tering, which in the case of bread relates to the degree of crystal-
development does not appear to be dependent on specific loaf linity of amylopectin, can be measured by NIR absorbance, so
volume (Fearn and Russell 1982). NIR can be used to follow the progress of bread staling (Wilson
Vol. 2, 2003—COMPREHENSIVE REVIEWS IN FOOD SCIENCE AND FOOD SAFETY 15
(Khoo and others 1975; Bechtel and others 1978), and pastes Biliaderis CG, Seneviratne HD. 1990. On the supermolecular structure and meta-
stability of glycerol monostearate-amylose complex. Carbohydr Polym 13:185.
(Fannon and BeMiller 1992 and references therein). Biliaderis CG, Tonogai JR. 1991. Influence of lipids on the thermal and mechanical
properties of concentrated starch gels. J Agric Food Chem 39:833.
Sensory/organoleptic tests Biliaderis CG, Vaughan DJ. 1987. Electron spin resonance studies of starch-water-
probe interactions. Carbohydr Polym 7:51-70.
Loss of flavor and aroma are among the most noticeable detri- Biliaderis CG, Page CM, Maurice TJ. 1986. Nonequilibrium melting of amylose-V
mental changes of bread upon staling. Reportedly, the decrease in complexes. Carbohydr Polym 6:269.
Biliaderis CG, Page CM, Slade L, Sirrett RR. 1985. Thermal behavior of amylose-lipid
the acceptability of bread over 5 d of storage is correlated with a complexes. Carbohydr Polym 5:367.
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Bowles LK. 1996. Amylolytic enzymes. In: Hebeda RE, Zobel H, editors. Baked
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Abstract: Staling of bread is cause of significant product waste in the world. We reviewed the literature of the last
10 y with the aim to give an up-to-date overview on processing/storage parameters, antistaling ingredients, sourdough
technology, and measurement methods of the staling phenomenon. Many researchers have been focusing their interest
on the selection of ingredients able to retard staling, mainly hydrocolloids, waxy wheat flours (WWF), and enzymes, but
different efforts have been made to understand the molecular basis of bread staling with the help of various measurement
methods. Results obtained confirm the central role of amylopectin retrogradation and water redistribution within the
different polymers in determining bread staling, but highlighted also the importance of other flour constituents, such as
proteins and nonstarch polysaccharides. Data obtained with thermal, spectroscopy, nuclear magnetic resonance, X-ray
crystallography, and colorimetry analysis have pointed out the need to encourage the use of one or more of these
techniques in order to better understand the mechanisms of staling. Results so far obtained have provided new insight on
bread staling, but the phenomenon has not been fully elucidated so far.
Keywords: bread, shelf life, staling
Introduction bread, waxy and high-amylose wheat (WT) starches and flours,
Bread stales and unfortunately it is a certainty and causes signif- sourdough, and analytical methodology (Table 1). More than 300
icant product waste all over the world (Collar and Rosell 2013). papers have been published in international peer-reviewed jour-
Staling results in loss of important sensory parameters of bread, like nals since 2003 on this topic, thus we attempted to collect the most
flavor and texture, and it is a consequence of a group of several important literature to give a new and up-to-date picture on bread
physical–chemical changes occurring during bread storage that staling. In particular, this review will focus on new information
lead mainly to an increase of crumb firmness and loss of freshness regarding the following aspects of bread staling: processing/storage
(Kulp and Ponte 1981; Gray and Bemiller 2003). Although the parameters, surface-active lipids, enzymes, carbohydrate ingredi-
staling mechanism has not been well established, the most impor- ents, flours and other major ingredients, as well as new measure-
tant causes responsible for this alteration are starch transformation, ment methods and sourdough technology. The review will take
starch–gluten interactions, and moisture redistribution (Schiraldi into consideration only papers dealing with WT bread and not
and Fessas 2001). with models such as diluted and concentrated starch pastes as well
Bread staling is being continuously studied and researchers have as gluten-free bread. In the case of papers dealing with the effect
been focusing their interest on mechanisms, factors, and mea- of different factors (such as storage temperature, ingredients, or
surement, thus a huge body of literature is available, including a ingredients of different origin), we use hierarchic considerations
number of reviews and book chapters dealing with the different to select the proper section of discussion. Moreover, the reader has
causes of bread staling and/or specific topics (Table 1). Most of the to refer to the literature previously reported and in particular to
reviews and book chapters do not cover all aspects dealing with the paper of Gray and Bemiller (2003) and others that will be cited
bread staling. A rather complete state of the art of molecular basis for more general information, molecular basis, and mechanisms of
and most of the factors influencing the quality of bread, as well bread staling. As a general rule, only papers not cited by specific
as of the main antistaling agents, have been, however, covered by or general reviews, which will be reported in the proper sections,
Gray and Bemiller (2003), while reviews published later focused are discussed in this review. However, papers already cited in re-
again only on specific aspects of bread staling, such as the influ- views, but not properly discussed with regard to bread staling will
ence of water, enzymes, frozen dough (FD), and partially baked be reviewed again.
C 2014 Institute of Food Technologists®
doi: 10.1111/1541-4337.12064 Vol. 13, 2014 r Comprehensive Reviews in Food Science and Food Safety 473
Bread staling review . . .
Table 1– Topics regarding bread staling covered by reviews or book WT flour by HBGB flours are more nutritious than those replaced
chapters. by CB flours and much more than regular WT flour breads pre-
Topic Review Book or book chapter serving the sensory acceptance and improving bread keepability
Enzymes Amos 1955; Haros Bowles 1996.
during storage. The high β-glucan content of barley flour (BM)
and others 2002; has been shown to help in reducing the starch crystallization, thus
van der Mareel and delaying significantly the staling rate of bread when used at the
others 2002; Butt
and others 2008; 20% level, even if it increased the firmness of fresh product (Gujral
Goesaert and and others 2003). Moreover, when BM was used together with
others 2009. wet gluten and ascorbic acid they reduced both initial firmness
Fibers Sivam and others
2010. and staling rate, especially when the higher level of the 3 addi-
Freezing and partial Rosell and Gomez tives was used. Purhagen and others (2008) proposed that water
baking 2007; Selomulyo had a greater effect on bread staling as assessed by texture anal-
and Zhou 2007; Yi
2008. ysis (TA), with respect to amylopectin retrogradation measured
Fundamental causes Kulp and Ponte 1981; with differential scanning calorimetry (DSC), when normal or
Le Meste and others heat-treated BMs were supplemented at 2% or 4% levels. In fact,
1992.
Hydrocolloids Izydorczyk and Dexter Milani and Maleki although the retrogradation enthalpy of supplemented breads was
2008; Kohajdová 2012. higher than control breads, the firmness values of barley loaves
and Karovičová
2009; Kohajdová were significantly lower during 7 d of storage at room tempera-
and others 2009. ture. However, the authors suggested that this effect could not be
Methodologies Karim and others Ponte and Faubion simply explained by the higher amounts of water in barley for-
2000; Chung and 1985; Ponte and
others 2003; Liu Ovadia 1996; mulations, but by differences in the water-binding ability of flour
and Scanlon 2004; Vodovotz and formulations with BM or soluble fibers. Staling rate was retarded
Choi and others others 2001. in laboratory-produced breads by using pressure-treated BM, as
2010.
Pentosans Hoseney 1984. well as waxy and pregelatinized waxy barley starch at the 3% level
Polyols Bhise and Kaur 2013. (Purhagen and others 2011b). The best results in retarding crumb
Proteins Davies 1986. firmness were found for pretreated and pregelatinized additives,
Sodium chloride Beck and others
2012a. with respect to the other formulations, including control bread,
Sourdough Arendt and others regardless of the storage time, even if a higher amylopectin ret-
2007; Chavan and
Chavan 2011. rogradation was revealed. The authors explained this result with
Starch Miyazaki and others Alsberg 1928; Slade the increased water retention during storage of substituted formu-
2006. and Levine 1987; lations. Unfortunately, they did not manage to retard staling when
Slade and Levine
1989; Hung and the pregelatinized additives were used in an industrial baking trial.
others 2006. Vittadini and Vodovotz (2003) used thermal analysis to assess
Staling Herz 1965; Zobel Alsberg 1936; that soy flour may have a role in modulating bread staling. Results
1973; Maga 1975; Chinachoti and
Knightly 1977; Vodovotz 2000; indicated that replacing up to 40% of soy flour in the bread formu-
D’Appolonia and Pateras 2007; lation caused a significant decrease in amylopectin recrystallization
Morad 1981; Cauvain and Young as well as promoted moisture retention during storage, with re-
Hoseney and Miller 2008.
1998; Schiraldi and spect to control bread, thus leading to decreased staling. Lodi and
Fessas 2001; Gray Vodovotz (2008) studied the effect of the partial substitution of
and Bemiller 2003.
Surface-active lipids and Knightly 1973; WT flour with soy flour and the addition of raw ground almonds
shortenings Stampfli and (5%). The incorporation of almond increased the loaf-specific vol-
Nersten 1995; ume of bread and reduced the crumb firmness changes over a 10-d
Kohajdová and
others (2009). storage period, if compared to bread obtained with only soy, even
Water Choi and others 2008. Cauvain and Young if no differences in amylopectin recrystallization rate or formation
2008. of amylose–lipid complexes were detected between the 2 formu-
lations. The authors postulated that the addition of almond to soy
flour probably resulted in a stronger interaction between proteins
Non-WT flours. It raises a great deal of recent interest that minor of WT and soy, favored by the high lipid content of almonds.
cereals, ancient crops, and pseudocereals, besides WT, constitute On the other hand, the bread produced with only soy staled at
highly nutrient-dense grains with feasible breadmaking applica- a lower rate than control bread, due to a better homogeneous
tions despite the poor viscoelasticity they exhibit when mixed water distribution, as revealed by different thermal determina-
with water. tions and by magnetic resonance imaging (MRI) (Lodi and others
Salehifar and Shahedi (2007) have confirmed earlier the ben- 2007a, 2007b).
eficial effects found by Zhang and others (1998) using oat flour Watanabe and others (2004) reported that substitution of WT
in reducing firmness of breads stored at room and chill temper- flour with powdered pre-germinated brown rice (PBGR) was
ature for up to 3 d, provided a maximum 20% oat flour sub- able to reduce the staling rate of bread stored for 3 d at room
stitution is accomplished, in order not to impart a strong bitter temperature, with respect to both control formula and bread sup-
taste. plemented with ungerminated brown rice (BR). The replacement
The ability of high β-glucan barley (HBGB) flour compared of 10% to 20% PBGR resulted in delayed staling with respect to
with regular commercial barley (CB) to make highly nutri- BR sample, while 10% PGBR slowed starch retrogradation, com-
tious WT-blended breads has been recently discussed (Collar and pared to control loaves, but supplementation of 30% PGBR ac-
Angioloni 2014a). Mixed breads obtained by 40% replacement of celerated bread hardening. According to the authors, 10% PGBR
474 Comprehensive Reviews in Food Science and Food Safety r Vol. 13, 2014
C 2014 Institute of Food Technologists®
Bread staling review . . .
addition enhanced softness of bread due to a certain amount of WWF substitution were softer up to 1 d in storage due to the
starch granules being gelatinized during PGBR production, while higher amount of water absorbed by the dough as well as the high
30% supplementation led to accelerated staling owing to the high moisture content in breadcrumbs. In a further paper, the same
water content needed to obtain dough. authors (Hung and others 2007b) by using 100% whole WWF
Mentes and others (2008) reported that substitution of WT flour managed to delay staling of whole waxy bread up to 3 d by adding
with ground flaxseed flour resulted in delayed staling of bread after 40000 U/g of cellulase, due to the particular pentosans present in
24 h in storage, with respect to control all–WT bread, as assessed the enzyme hydrolysate. Moreover, they obtained white WWF by
by a mechanical penetration test, but the authors did not give any removing the bran and germ, and the resulting breadcrumbs kept
explanation of the probable causes. The best result was obtained softer for 5 d, with respect to breadcrumbs from both the whole
by using 15% flaxseed flour. regular and whole waxy WT, probably as a result of the enrich-
Wu and others (2009) studied the effect of potato paste sub- ment of the amylopectin fraction of the white WWF. Park and
stitution at 5% to 30% on hardness evolution of bread during a Baik (2007) made a comparative test with WT genotypes of wild
3-d storage period and found that staling decreased in 1-d stored type, partial waxy, and waxy starch, in order to study the influence
samples obtained with 5% to 20% potato paste, with respect to of starch amylose content on French bread performance of WT
control breads, and they associated this with the differences in flour. Their study evidenced that WT flours with reduced starch
water-binding capacities of potato paste and with interaction with amylose content allowed the production of breads with better re-
starch, thus affecting starch retrogradation. tained crumb moisture and delayed staling up to 48 h of storage,
Begum and others (2010) evidenced that bread obtained with probably because the greater crumb moisture resulted in a delay in
the use of 10% fermented cassava flour or 10% soy-fortified cassava amylopectin retrogradation, even if DSC analysis did not evidence
flour was softer after 3 d at room temperature, with respect to WT significant differences in enthalpy values of the various WT geno-
bread (Note: the authors did not make an explanation for this types with different amylose content. Slowing the migration of
result and did not report the amount of soy used to fortify cassava water from the gluten phase to the starch phase by WWF (5% to
flour). 30%) has been hypothesized as the cause of diminution of firmness
In a recent paper (Angioloni and Collar 2011), the suitability evolution, as determined with compression analysis (Mouliney and
of associated mixtures of minor/ancient cereals (rye, oat, Kamut R
others 2011).
WT, spelt WT) and pseudocereals (buckwheat) to obtain baked The low amylose content of flours obtained from 2 new Japanese
goods with high characteristics was assessed in multigrain WT WT varieties was related to reduced staling of bread, especially in
flour highly replaced matrices. A quaternary blend of oat, rye, the first 48 h of storage at 20 °C, with respect to samples obtained
buckwheat, and common WT flours (20:20:20:40 w/w/w/w) with 2 representative bread WT classes that are N. 1 Canada west-
without any additive and/or technological aid in the formulation ern red spring and hard red winter (Ito and others 2007). DSC
was proposed to make highly nutritious baked goods meeting data of enthalpy and X-ray patterns evidenced a slow retrogra-
sensory standards and exhibiting a low staling rate during ageing. dation of starch gel in the bread obtained with the new varieties,
The quality profile of binary mixtures of oat–WT (60:40 w/w), thus accounting for their softer texture that resulted in softness and
millet–WT (40:60 w/w), and sorghum–WT (40:60 w/w) was sig- high cohesiveness of the loaves. Apparently different results were
nificantly improved in presence of some additives in terms of keep- found when replacing hard WT flours with 15% to 45% with two
ability during storage, mainly for oat–WT blends, which stale at a hard WWF (Garimella Purna and others 2011). In fact, substi-
similar rate or even at lower rate than 100% WT breads (Angioloni tution led to softer bread, but only at day 1 after baking, while
and Collar 2013). Dilution up to 20% of the basic rye/WT flour staling was not retarded during storage. The combination of less
blend by accumulative addition of amaranth, buckwheat, quinoa, amylose and more soluble starch from amylopectin characterizing
and teff flours (5% single flour) did positively impact both bread WWF could have resulted in a soft crumb structure on day 1 after
keeping behavior during aging, and nutritional characteristics of baking, while after 7 d the bread was as firm as the control, due
mixed bread matrices (Collar and Angioloni 2014b). to a similar content of soluble starch, thus confirming a previous
Waxy WT flours (WWF). Most of the research work on flour has study (Ghiasi and others 1984).
been focused, however, on the use of WWF, because, due to its lack Yi and others (2009) studied the effect of partial WWF substi-
of amylose, WWF can reduce the initial phase of retrogradation tution on staling of bread made from FD. They found that when
(Graybosch 1998). A comprehensive review on the production modulating WWF and water amounts it was possible to reduce the
and characteristics of WWF and waxy wheat starch (WWS) and staling rate, with respect to control formulations. The best com-
their application for food processing is that of Hung and others bination was 45% WWF replacement and 65% water. By using
(2006). pulsed hydrogen-1 nuclear magnetic resonance (1 H NMR) they
Baik and others (2003) suggested that the increased starch ret- concluded that bread with higher WWF content held more water
rogradation of bread crumb, as assessed by DSC, may not be the and limited the movement of water from one domain to another.
cause of retarded staling during a period of 7 d in storage at 4 °C Very recently, Lafaye and others (2013) obtained bread using
in bread obtained with double-null partial WWF, with respect to waxy durum flour and concluded that this flour acted as a unique
bread produced with hard red spring WT flours. They proposed bread softener. The authors did not make any additional analysis
that the low amylose and high-protein contents of the waxy lines in order to suggest a satisfactory explanation of the antistaling
were beneficial in retarding the increase in hardness. Peng and effect of this flour, however provided a well-described picture of
others (2009) reported that the use of 15% WWF combined with the possible causes leading to the beneficial effect of waxy flour
2 other WT flours was the optimal solution for retarding staling supplementation by summarizing literature results.
up to 6 d without impairing bread quality, as revealed by sensory
analysis, if compared with the control. Data from Hung and others Carbohydrates
(2007a) gave evidence of the relationship between the use of whole A consistent research activity has been carried out during the
WWF and delayed staling. Breads made with 30% and 50% whole last decade on the role of carbohydrate ingredients in reducing
C 2014 Institute of Food Technologists® Vol. 13, 2014 r Comprehensive Reviews in Food Science and Food Safety 475
Bread staling review . . .
Table 2– Main hydrocolloids proposed during the last decade for staling reduction.
bread staling. Hydrocolloids, modified starches, dextrins, and mal- Ghanbari and Farmani (2013) who revealed a significant antistaling
tooligosaccharides and other fibers will be covered in this section. effect of K, especially when supplemented at 0.5%. Mandala and
Hydrocolloids. The antistaling effect of hydrocolloids (Table 2) Sotirakoglou (2005) suggested that the use of XG and GG in fresh
has been extensively studied and attributed to controlling and or microwave-heated bread after frozen storage was able to retain
maintaining the moisture content, stabilizing the dough, and in- water in the crumb and, consequently, moisture migration to the
fluencing the crust structure (Davidou and others 1996; Collar and crust, thus resulting in the crust to fail at greater deformation,
others 1999; Rojas and others 1999; Mandala and Sotirakoglou that is, the samples were less stiff. XG used at low concentrations,
2005; Mandala and others 2007; Rosell and Gomez 2007). Some on the other hand, improved the crumb viscoelastic properties on
interesting reviews focused on molecular structure, physicochem- defrosted and microwave-heated samples, probably by hindering
ical properties, and uses in food products of the whole class the deteriorating effects and avoiding the development of a spongy
of hydrocolloids as bread improvers (Kohajdová and Karovičová structure during frozen storage, as suggested by Ferrero and oth-
2009) and more specifically of barley β-glucans and arabinoxylans ers (1993). Moreover, XG has been addressed to retard amylose
(Izydorczyk and Dexter 2008). A book chapter by Milani and retrogradation, due to reduced amylose–amylose interactions. In
Maleki (2012) gives a classification of hydrocolloids and of their 2 separate papers the effect of 4 different hydrocolloids was stud-
functions, according to Hollingworth (2010). ied, namely XG, GG, locust bean gum (LBG), and HPMC on
The use of DSC allowed to establish that hydroxypropyl methyl- staling retardation of dough bread (DB), par-baked (PB) bread,
cellulose (HPMC) and k-carragenan (K) decreased the retrograda- and full-baked (FB) breads stored at chilling (Mandala and oth-
tion enthalpy of amylopectin, thus retarding staling of part-baked ers 2007) or frozen temperature (Mandala and others 2008) and
breads produced with an interrupted baking process and frozen finally re-baked (DB and PB). The crust puncture test and re-
storage (Barcenas and others 2003). The latter results were, in laxation test of the crumb revealed that XG addition resulted in
part, in contrast to what was reported previously by Sharadanant a significantly less firm crust on PB and FB breads after chilling
and Khan (2003) who found a detrimental effect on bread firm- storage, with respect to the other samples. X had also the more
ness evolution during storage of K-supplemented breads. In a later evident effects on crumb viscoelastic properties, as revealed by re-
paper, Barcenas and Rosell (2005) gave a more detailed explana- laxation tests, as it gave PB breads with an elastic crumb, DB with
tion of the possible cause of the antistaling effect of HPMC. The a more viscous crumb, and FB breads with an even more viscous
authors, in fact, determined the microstructure of bread crumb by crumb (Mandala and others 2007). In the case of frozen sam-
cryo-SEM and found that HPMC use resulted in gas cells with ples (Mandala and others 2008), XG supplementation was able to
a more continuous surface and a thicker appearance, with respect give a softer plastic crust, but only in PB breads, with respect
to the control. Thus, the presence of HPMC enfolded the other to control and other supplemented samples, probably due to the
bread constituents, with a consequent hindering of their interac- thickening effect on the crumb walls associated with the air spaces
tions and avoided some of the processes involved in bread staling. that resulted in a less rigid structure. Finally, the addition of XG to
The HPMC was suggested as the best antistaling ingredient also formulations allowed PB and FB breads to have a more elastic
for Lavash flat bread made with 2 different WT flours and stored crumb when compared to the other samples, thus revealing that
for 48 h (Tavakolipour and Kalbasi-Ashtari 2007). Similar results this hydrocolloid is more efficient against crumb deterioration in
on another flat bread, the Barbari, have recently been reported an FB product than in the DB, and highlighting a very different
by Maleki and others (2012) who found that hydrocolloids other behavior from that found during chilling storage (Mandala and
than HPMC, namely guar gum (GG), xanthan gum (XG), and others 2007), in which FB breads presented a complete viscous
carboxymethylcellulose (CMC), reduced staling of bread up to 5 and deteriorated crumb when hydrocolloids were used. Shittu and
d, due to the limitation of water mobility that influenced the gela- others (2009) reported that increasing the dosage of XG up to 2%
tinization process by decreasing the H, that was also reported by resulted in a major hindrance of gluten–starch interaction in the
476 Comprehensive Reviews in Food Science and Food Safety r Vol. 13, 2014
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Bread staling review . . .
presence of hydrocolloid molecules, thus conferring a significantly at 2.5% into a bread formulation reduced crumb staling after 1 d in
higher softness to fresh composite cassava–WT bread. They also storage, as detected by TA, but they did not formulate any expla-
reported that crumb hardening and moisture loss followed a linear nation of the causes (Note: the discussion of data on firming is not
sequence up to the 1% XG level, which, thus, was proposed as the exhaustive, as the authors neither explained the rate of staling, nor
optimum concentration to reduce both phenomena, even if the highlighted that there were no significant differences in firmness
2% XG level best estimated the crumb firming rate. Shalini and between control and BGF-supplemented bread). The BGF gave,
Laxmi (2007) investigated the effect of 4 different hydrocolloids, moreover, breads with lower volume, confirming previous results
HPMC, GG, K, and CMC on textural characteristics of Indian (Gill and others 2002). Jacobs and others (2008) gave interesting
chapatti bread stored at ambient or chilling temperature and evi- new knowledge about the influence of bread production on bread
denced that 0.75% w/w supplementation of GG gave the softest quality when fiber-rich fractions (FRF), enriched β-glucans, and
bread and decreased the loss of extensibility up to 2 d in storage at arabinoxylans from hull-less barley were used. They, while con-
both temperatures, with respect to the control. The authors sug- firming the results of Symons and Brenan (2004), found that sup-
gested that GG has a softening effect, probably by an inhibition of plementation of FRF (12% on flour basis, corresponding to 2.5 g
the amylopectin retrogradation, prevention of water release, and of arabinoxylans and β-glucans per 100 g of flour) and Xyn within
polymer aggregation during refrigeration, as well as interference the sponge-and-dough (SAD) baking method, improved the loaf
during interchain–amylose association. In a further paper, Shalini volume, appearance, and crumb structure and resulted in crumb
(2009) gave more explanations on the effects of GG on staling hardness and staling rate similar to that of the control bread, while
parameters and found that moisture, water-soluble starch, and in other baking methods (Canadian short process, remix-to-peak)
vitro digestibility enzyme contents in GG-incorporated chapatti gave negative results (Note: the main part of the paper deals with
were higher than in the control chapatti at both storage temper- a comparison of the 3 baking methods by using a 20% on flour
atures. Smitha and others (2008), on the other hand, working basis supplementation and the authors concluded that the qual-
with another flat bread, the unleavened Indian parotta, found that ity of the 20% FRF-enriched SAD bread was equal to or better
supplementation of hydrocolloids (gum arabic, GG, XG, CMC, than the remix-to-peak bread, but they neither presented a statis-
and HPMC) resulted in delayed staling 8 h after baking, with tical comparison between data of the 2 baking methods, nor did
respect to nonsupplemented breads. HPMC gave the best results they explain why they evaluated the impact of lower FRF addition
in terms of reduction of hardness, while XG was judged by pan- only with the SAD method). Skendi and others (2010) studied the
elists as the best for preserving sensory attributes like softness and supplementation of 2 WT flours differing in bread making qual-
chewiness. Angioloni and Collar (2009a) proposed the viability of ity (poor and good) with two different-molecular-weight barley
LBG and CMC blended with oligosaccharides, at a medium-high β-glucan isolates (at 0%, 0.2%, 0.6%, 1.0%, and 1.4% w/w on
substitution level, as very valuable sources of dietary fiber (DF) a flour dry weight basis) and found that the crumb hardness of
for the baked goods with both “healthy” characteristics and ex- β-glucan–supplemented breads, measured after 24 h of storage,
tended shelf-life, due to reduced staling. These conclusions were decreased with its increasing level up to reaching a minimum, and
drawn after modeling the crumb firming kinetics parameters ob- then with a reverse trend, however the values were always lower
tained during storage with the Avrami equation. Moreover, good than the control bread, if we ignore one sample. Moreover, the
relationships between the main parameters obtained with the dif- antistaling effect was more pronounced up to 8 d in storage when
ferent physical analyses (small dynamic and large static deformation the higher-molecular-weight β-glucan isolate was used in both
methods, viscometric pattern, and image analyses) performed on flour types. The authors proposed that the beneficial effects found
raw materials and intermediate and final products were found. could be ascribed either to the higher water retention capacity
The effect of sodium alginate (ALG) and konjac glucomannan and a possible inhibition of the amylopectin retrogradation of β-
(KGM) supplementation at 0.2% and 0.8% w/w flour basis was glucan, as already suggested (Biliaderis and others 1995), or to the
studied in terms of staling behavior of Chinese steamed bread by increase of the total area of gas cells.
Sim and others (2011) who reported that the higher supplemen- An increase in bread firmness with respect to control WT for-
tation dose of both hydrocolloids resulted in a significantly lower mulation was, on the other hand, found by Hager and others
staling rate up to 4 d, with respect to the control bread, probably (2011) after addition of oat β-glucan, suggesting that this increase
because of the hindering effect of gums on macromolecular en- in hardness might be ascribed to the increased water-binding ca-
tanglements thus causing starch recrystallization delay. Wang and pacity of the polysaccharide, thus hindering the development of
others (2006) studied the effect of gluten hydrolysate (GHP)/λ- the gluten network. They also evidenced a consistent increase in
carrageenan (λC) ratio on the increase in the bread crumb firm- staling after the addition of the fat replacer inulin, thus confirm-
ness during storage and proposed that the changes occurring in ing previous results (Wang and others 2002; O’Brien and others
the amorphous part of the starch, when a concentration of 0.5% 2003; Poinot and others 2010) and in part in agreement with
GHP/γ C was used in the product formulation, thus significantly the study of Peressini and Sensidoni (2009) who used 2 com-
delaying bread staling. mercial inulin products, with lower (ST) and higher (HP) degrees
The use of hemicelluloses has been the topic of different studies of polymerization, to supplement 3 different WT flours, moder-
during the last 10 y. A penetrometric test revealed that supple- ately strong (MS) and weak (W), and found that the ST inulin
mentation of 0.3%, 0.5%, or 0.7% hemicelluloses (extracted from addition to MS flour significantly increased the volume and low-
buckwheat) increased the penetration depth of the crumb after 72 ered bread firmness, with respect to the control. The authors
h of storage at 30 °C, thus delaying crumb hardening, and resulted hypothesized that a delayed starch gelatinization during baking,
in bread with a higher specific volume than the control during a due to the presence of 12% solutes, and the significant reduc-
3-d storage period. The best results were in the order 0.5% > 0.3% tion of dough water absorption of ST inulin, may explain this
> 0.7% both for hardness and volume (P = 0.01) (Hromádková result. The beneficial effect of inulin gel at 2.5% flour basis on
and others 2007). Symons and Brennan (2004) reported that a β- increasing the loaf volume and maintaining the hardness value,
glucan-rich fraction (BGF) extracted from barley and incorporated with respect to a control bread, was also reported by O’Brien
C 2014 Institute of Food Technologists® Vol. 13, 2014 r Comprehensive Reviews in Food Science and Food Safety 477
Bread staling review . . .
and others (2003). In a very recent paper the antistaling effect of staling). In a further paper, they showed that supplementation
of substitution of WT flour with BM (28%, 56%, and 84%) or of ascorbic acid to chitosan-enriched bread resulted in a more pro-
β-glucan (1.5%, 3.0%, and 4.5%) on chapatti bread was assessed by nounced decrease of water content during baking in fresh bread
DSC (Sharma and Gujral 2014). Storage at 4 °C for 24 h induced compared to the control bread (Kerch and others 2012b).
retrogradation in baked chapatties, as revealed by the increase in Modified and damaged starches. The use of modified starches
enthalpy of melting (H), but it was concomitantly reduced up for retarding staling has been suggested since the 1990s, for their
to 44% or 64% when β-glucan or BM was used, respectively. ability to influence amylopectin crystallization (Inagaki and Seib
The authors proposed that BM supplementation increased the 1992; Yook and others 1993; Toufeili and others 1999). Due to
levels of soluble as well as insoluble DF, with an increased water the fact, however, that other linear fractions of starch may affect
absorption and change of the nature of the starch and protein, retrogradation, an increased interest has been registered on cross-
thus preventing better the staling of chapatties, with respect to linked starches, due to their ability to increase the gelatinization
loaves obtained with β-glucan alone, as suggested by Purhagen and temperature, setback viscosity, and decrease the transition enthalpy
others (2012). of gelatinization (Zheng and others 1999; Woo and Seib 2002). A
The use of pectin slowed crumb hardening in bread that was well-focused review on this topic has been published by Miyazaki
part-baked and stored at chilling (PB) or subzero (PBF) temper- and others (2006).
atures for variable times (Rosell and Santos 2010). The authors According to Leon and others (2006), the content of damaged
also revealed that PBF pectin-supplemented breads showed a sim- starch directly influences bread staling through the increase of
ilar hardening trend, with respect to their conventionally baked amylopectin recrystallization, as detected by DSC analysis. The
counterparts, as also demonstrated by using the Avrami equation. authors concluded that the limited use of damaged starch is a key
Correa and others (2012) reported that the incorporation of high- factor to control the quality of fresh bread and of its shelf-life,
methoxyl pectin at 1% or 2% resulted in protection with respect in contrast to what was reported earlier by Tipples (1969). In a
to staling, especially when salt was used in the formulation, as paper of Miyazaki and others (2008), chemically modified tapioca
it reduced the hardness values with respect to the control sam- starches (MTS), but with different degrees of modification, have
ple, as well as maintaining the chewiness. They proposed that the been used to retard staling in breads obtained from FD, which
improved specific volume of high-methoxyl pectin-supplemented was subjected to one freeze–thaw cycle and 1-wk frozen storage.
bread, which gave both a more cohesive and more resilient crumb Highly MTS retarded significantly the increase in firmness during
with a different alveolus structure, was the main reason for retarded 3 d of storage, thus confirming the results of previous papers, due
staling. to the slow retrograding rate of amylopectin.
A certain interest during the last years has been focused also on Dextrins and maltooligosaccharides. Dextrins are the product
an animal hydrocolloid, chitosan, a nonbranched linear homopoly- of starch hydrolysis and, since bread staling has been attributed
mer obtained from shrimp and other crustacean shells. Chitosan is partly to its retrogradation, shortening the starch chain length, as
a water-soluble cationic polyelectrolyte, while most of the other obtained with particular α-amylases, results in reducing the rate of
polysaccharides are neutral or negatively charged at acid pH. In staling.
a first paper of Kerch and others (2008), addition of 2% chitosan Miyazaki and others (2004), using DSC, found that among
resulted in increased staling rate of bread, and the author, through 6 different dextrins (dextrose equivalent 3 to 40) used at 20%,
DSC analysis and SEM, suggested an increase in water migration those with low molecular weight (DE 19, 25, and 40) at 2.5%
rate from crumb to crust and in dehydration rate both for starch of substitution retarded retrogradation, as revealed by the H of
and gluten and a prevention of amylose–lipid complexation in retrograded amylopectin, but did not delay staling during 3 d
breads supplemented with chitosan. of storage. They postulated that the antistaling mechanism fol-
In a following paper, Kerch and others (2010) proposed and lowing addition of dextrin differed from the retarding effect of
analyzed, with the aid of mechanical and DSC measurements, dextrin produced by α-amylase, as already reported (Akers and
the main possible mechanisms leading to staling in breads ob- Hoseney 1994; Morgan and others 1997). They also highlighted
tained with supplementation of different chitosan and chitosan that retrogradation is not related to water mobility in crumbs, as as-
oligosaccharides. They confirmed that staling was the result of sessed by the determination of water activity. An interesting study
2 independent processes, the first during the first 2 d of stor- involving the use of texture profile analysis (TPA), X-ray diffrac-
age depended on changes in the organization of starch polymer tion (XRD), and DSC reported that the use of β-cyclodextrin
chains, and later on it was caused by loss of water by gluten. (β-CD) resulted in retardation of bread staling during 35 d of
They suggested also that chitosan increased the firming rate dur- storage at 4 °C, as changes of some TPA indexes (hardness, co-
ing the 1st stage due to its ability to bind lipids and prevent hesiveness, and springiness) were reduced (Tian and others 2009).
amylase–lipid complexation, while in the 2nd stage it was en- Data on hardness were fitted with the Avrami equation that evi-
hanced dehydration of gluten due to its water-binding ability. In denced a significant reduction of the rate constant (k), while in-
their work, however, they found that both chitosan oligosaccha- creasing the Avrami exponent, thus suggesting a retarded crumb-
rides and low-molecular-weight chitosan decreased significantly firming kinetic for β-CD-supplemented bread. Moreover, data
the staling rate, if compared to middle-molecular-weight chitosan, of XRD showed a delay in changes of crystalline patterns occur-
and they hypothesized that low-molecular-weight substances in- ring in crust and crumb and this retardation was attributed to a
hibited crosslink formations between starch granules and protein complex amylose–lipid β-CD, as observed by DSC, that resulted
fibrils which, in turn, are responsible for staling. Later on, Kerch in transformation of nucleation type and lowered rate of bread
and others (2012a) demonstrated with DSC that when chitosan staling.
was used in bread production by the straight-dough or the SAD Jakob and others (2012), studied for the first time, the beneficial
method it accelerated or slowed down the decrease of bound water effects of different fructans produced by acetic acid bacteria on the
content during the 1st stage of staling, respectively, thus delaying texture of bread. Out of 21 strains tested, 4 of them were able to
or accelerating staling during the first 2 to 3 d of storage (1st stage produce high amounts of exopolysaccharides (EPS), as detected
478 Comprehensive Reviews in Food Science and Food Safety r Vol. 13, 2014
C 2014 Institute of Food Technologists®
Bread staling review . . .
by HPLC analysis, which elicited, when supplemented at 1% to polysaccharide, which was able to reduce the slope of the stal-
2% of flour basis, significantly the increase in bread volume and ing rate up to 9 times, with respect to the control. The authors
retarded the hardness increase of crumb up to 1 wk of storage, also found that the magnitude of bread staling retardation strongly
the highest differences being observed after addition of 2% sugar depended on the type of WT flour used.
polymer from Neoasaia chiangmaiensis. The authors proposed that Addition of butternut fiber at 10 g/kg of flour decreased the
the functional properties of the tested EPS were due to their hy- staling rate of bread after 7 d of storage, as measured by compress-
drocolloid character, allowing a high water retention, and due to ibility, DSC, and digital image analysis (Pla and others 2013). The
interactions between polysaccharides and other dough compo- authors clearly showed that fiber extracted from the peel resulted
nents like gluten and starch, thus influencing the final structure of in a drastic reduction of the firmness value, suggesting a retarda-
the baked product. They, moreover, compared effects of EPS to tion of amylopectin retrogradation, more air occluded (cell area
HPMC. 100/total area), same number of particles (alveolus or gas cells) per
Other fibers. In this section the effect of DF other than pre- square centimeter, and higher mean size of particles, with respect
viously defined hydrocolloids and dextrins on bread staling will to the control bread. The authors concluded that the particular
be reviewed. According to the Codex Alimentarius Commis- composition of fiber extracted from peel, that is presence of lignin,
sion (2009), DF are “Carbohydrate polymers with more than 10 less-branched pectin chains, and significant higher protein content
monomeric units, which are not hydrolyzed by the endogenous than the other butternut fiber used, may have accounted for the
enzymes in the small intestine of humans” (ALINORM 09/32/26 results obtained.
2009). Recently dough properties of bread enriched with DF have
been reviewed and the reader is redirected to this paper for the Lipids and shortenings
aspects dealing with the interaction of this component in dough The role of surface-active lipids and shortenings has been well
development and bread baking (Sivam and others 2010). Fibers described by Gray and Bemiller (2003), and later on by Kohajdová
investigated during this last decade are of cereal or noncereal ori- and others (2009), thus we will report briefly the new knowledge
gin. not or only partially covered by these 2 reviews focused mainly on
Maeda and Morita (2003) proposed the polishing of soft WT the use of mono or diacylglycerols alone or esterified (DATEM).
grain from the outer layer in increments of 10% of total weight to Collar (2003) suggested that individual and/or binary sup-
obtain flours with a high content of pentosans and damaged starch. plementation of fat-monoglycerides (MGL) and sodium stearoyl
In particular, both water-soluble (WSP) and water-insoluble pen- lactylate (SSL) to bread dough positively influenced the level of
tosans (WISP) from the inner part of the WT grain were added to the pasting parameters assessed by rapid visco-analysis (RVA) (peak
the conventional flour and their effects on loaf volume and bread viscosity, pasting temperature, and setback during cooling) that are
staling were assessed. The results indicate that both pentosans gave associated with a significant delay in bread firming. Moreover, she
an increase in loaf volume and a significant decrease in staling up does not recommend binary use of MGL/CMC and SSL/CMC,
to 3 d in storage, with respect to the control bread. The authors as the antagonistic effects of the pair gum/surfactant resulted in a
presumed that the high viscous and gelling properties of WSP may nullification of the benefits exerted by the individual emulsifiers.
improve the strength of gluten and the retention of gas generated Ribotta and others (2004a) evidenced the beneficial effect of
in the dough. DATEM on retarding crumb firming at 4 and 20 °C aging tem-
Mandala and others (2009) studied the effect of different ingre- perature of bread from both nonfrozen and FD, and they supposed
dients (hydrocolloids, polydextrose, oat flour, inulin, and commer- that the formation of complexes with amylose and amylopectin
cial shortening) on crust firmness and crumb elasticity of breads inhibited the staling phenomenon. Sawa and others (2009) studied
obtained after thawing and baking of FD (at subzero temperatures the effects of a wide range of purified saturated and unsaturated
for 1 wk) and PB breads, and found that inulin was the best of MGL at different concentrations on the crumb firmness evolu-
them in reducing bread crust firmness, probably due to a better tion during bread storage and reported that the use of C16:0 and
moisture redistribution, even if fresh sample had the firmer crust. C18:0 and cis- and trans- C18:1 resulted both in a lower crumb
Gomez and others (2003) found that the use of fibers of differ- firmness, even if depending on the baking process used, and in
ent origin (cellulose, cocoa, coffee, pea, orange, and WT), while delayed bread staling, when compared to control bread. They sug-
increasing the crumb firmness of fresh bread with respect to the gested the interaction of MGL with amylose and amylopectin
control, reduced its evolution during 3 d of storage, and they as the main cause of the obtained results. Manzocco and others
postulated that this effect may be attributed to the already demon- (2012) proposed that a particular system morphology, as assessed
strated water-binding capacity of fiber, which in turn reduced wa- by proton density/mobility using MRI, was generated in bread in
ter loss during storage, as well as the probable interaction between which palm oil was replaced with a MGL–sunflower oil–water gel.
fiber and starch, resulting in the delay of starch retrogradation. The The morphology change resulted in an 81% reduction in bread
best effect in delaying the bread staling was noticed after 2 d by fat content as well as in a delay in bread staling during storage.
using a short-length WT fiber. The incorporation of the gel resulted, in fact, in a reduced proton
Collar and others (2009) found a positive effect on reducing density/mobility in comparison with standard formulation, thus it
staling rate during 16 d of storage of breads enriched with 2 kinds was concluded that the physical architecture of the lipids used in
of cocoa fiber, as assessed by hardness and chewiness fitted with the formulation could contribute to modulate the retrogradation
the Avrami equation, when increasing the dose of addition up rate. Smith and Johansson (2004) reported that the increase of solid
to 6%, especially when the formulation was supplemented with fat of a shortening containing fully hydrogenated soybean oil was
alkalinized cocoa-soluble fiber, while over-dosage resulted in a able to delay bread staling and they suggested that saturated tria-
staling rate similar to that of the control breads. cylglycerols acted in a similar way as saturated monoacylglycerols,
Zhou and others (2009) correlated the reduction of starch that was an interaction with amylopectin. Mnif and others (2012)
retrogradation after X-ray measurements and application of the proposed a new biosurfactant obtained by Bacillus subtilis as anti-
Avrami model with increasing levels, from 1% to 5%, of tea staling agent and compared it to soy lecithin. The bioemulsifier
C 2014 Institute of Food Technologists® Vol. 13, 2014 r Comprehensive Reviews in Food Science and Food Safety 479
Bread staling review . . .
supplementation significantly reduced the bread staling over an Chin and others (2012) focused their work also on crust behav-
8-d period, depending on its amount, and the maximum decrease ior following different glazing applications. Glazing with corn-
of staling rate was obtained with a 0.05% biosurfactant addition, starch, skim milk, and egg white were able to reduce the rate of
which was also the dose that resulted in the highest bread-specific moisture loss in bread crumb during 6 d of storage, thus reducing
volume. The authors suggested that the slower firming may be the staling rate of glazed bread, with respect to the control. More-
ascribed to the capacity of the emulsifiers to form a complex over, glazing resulted in an increase in crust firmness, although the
starch-emulsifier, which in turn delayed WT starch crystalliza- moisture content of the crust increased, probably because the rate
tion. Additionally, the biosurfactant reduced the susceptibility to of moisture migration from crust to the surrounding atmosphere
microbial growth during bread storage. could be lower with respect to that from the crumb to the crust
region.
Minor Ingredients Affecting Bread Staling Sodium chloride impact on bread staling has recently been well
The functional effects on fermentation and bread baking of reviewed and ascribed mainly to the increased gas retention effect
whey protein and casein have been reported by Erdoghu-Arnozcki of dough with NaCl that allows an increase in crumb porosity
and others (1996). Casein and whey, together with ALG and K and a consequent decrease in crumb firmness (Beck and others
were used in an attempt to improve the quality of FD, specifically 2012a). The retrogradation effect ascribed to Na+ inclusion in
to retard its quality loss during freezing time and after 3 freeze– starch molecules during storage of bread has been suggested as
thaw cycles (Yun and Eun 2006). Bread made with milk proteins delaying staling (Beck and others 2012b). In particular, a decrease
and hydrocolloids were softer after 4 d in storage, with respect to in bread staling following the decrease in NaCl levels was shown.
control bread, probably because of better moisture retention and Furthermore, a linear relationship between rheofermentometer
improved emulsification of these ingredients. Similar results were data, bread volume, and crumb firmness was demonstrated, thus
obtained in a later paper of Shon and others (2009). suggesting that the quality of bread could be predicted by gas
Addition of juices to WT bread formulations have been pro- release measurement.
posed to ameliorate its nutritional profile (Batu 2005), as sweet-
eners and color enhancers, and to increase volume and extend Enzymes
shelf-life (Matz 1989). Lasekan and others (2011) postulated that The role of enzymes on bread staling has been one of the
the high concentration of monosaccharide of the pineapple juice preferred topics during this last decade and along with quite recent
concentrate used at a 1.5% level in the formulation of white bread reviews (Haros and others 2002; Butt and others 2008; Goesaert
interfered with protein–starch interaction and delayed staling, but and others 2009), an important number of papers have appeared,
only after 1 d of storage. Sabanis and others (2009) studied the which will be discussed. Apart from the effects of amylases, an
effect of supplementation (at 50% level sucrose substitution) of increasing interest in transglutaminase, a protein modifier enzyme
2 types of raisin juices, concentrated and dried on evolution of and other nonstarch polysaccharide-modifying enzymes has been
crumb firmness of bread obtained with both bread WT and du- recorded.
rum WT. The dried juice decreased the WT starch gel rigid-
ity and retrogradation for the presence of glucose and fructose, α-Amylases and transferases
thus resulting in reduced staling after 2 d of storage, with re- The action of α-amylases in reducing bread staling has been the
spect to control loaves, especially when durum WT flour was topic of numerous studies (Gray and Bemiller 2003), and different
used. ways of action have been proposed. The paper of Goesaert and
Tomato pomace has been suggested as a good source of hydro- others (2009) provided new knowledge on the α-amylase mode
colloids and was thus proposed (0%, 1%, 3%, 5%, and 7%) for flat of action and its antistaling activity. In particular, they found that
bread production by Majzoobi and others (2011b), who detected the maltogenic α-amylase from Bacillus stearothermophilus degraded
delayed bread staling up to 4 d of storage at 25 °C in tomato significantly the outer amylopectin branches, thus producing amy-
pomace–supplemented bread, with respect to control sample, due lopectin chains that are too short to crystallize. The result was the
to the concomitant increase in volume and moisture content and prevention of a “permanent” (based on amylopectin crystallites
decreased starch retrogradation. junction zones) amylopectin network, thus staling was delayed.
Surface coating treatments have been patented for improving Maeda and others (2003) proposed that a particular thermostable
the quality of bakery products (Lang and others 1987; Lonergan mutant, α-amylase (M77), purified from Bacillus amyloliquefaciens
1999; Hahn and others 2001; Jacobson 2003; Casper and oth- F increased the specific volume of the bread and improved the
ers 2006, 2007). The main advantages proposed for glazing were softness of bread crumb, when compared to the commercial exo-
the improvement of flavor and appearance. The moisture barrier type α-amylase Novamyl (NM). They also showed that softness
exerted at the surface of baked products allows retaining and aid- evolution of breadcrumb during storage was not correlated with
ing dough expansion during baking, thus resulting in a reduction thermostability. Rao and Satyanarayana (2007) found that the ad-
of surface defects, improvement of color, and higher baked vol- dition of α-amylases produced by Geobacillus thermoleovorans to
ume. Recently, however, other beneficial effects of glazing have WT flour improved the fermentation rate and decreased the vis-
been described. Jahromi and others (2012) studied the effect of cosity of dough, while increasing the volume and texture of bread,
different glazing treatments, including natural substances, polyol, moreover, it also increased its shelf-life by retarding staling, with
sugar, and hydrocolloids on the staling rate of breads stored up to respect to control sample, but they did not give any explanation of
12 d. Increased moisture and reduction of water movement have this beneficial effect. Jones and others (2008) managed to develop
been addressed as the main causes of delayed staling by differ- a new maltogenic α-amylase from Bacillus sp. TS-25, formerly B.
ent glazing ingredients, mainly, water, egg yolk, propylene glycol, stearothermophilus, which increased thermal stability and the possi-
and starch, while at intermediate storage periods (2, 5, and 8 d) bility to work at acidic pH values that are typical of sourdough and
also other glazing substances significantly retarded the increase in rye breads. Kim and others (2006) reported that the addition of
crumb firmness, with respect to control bread. a fungal α-amylase to polished flour resulted in an improvement
480 Comprehensive Reviews in Food Science and Food Safety r Vol. 13, 2014
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Bread staling review . . .
of gas cell distribution and softness of breadcrumbs and delayed to the formation of amylase–lipid complexes, both revealed by
staling, without lowering the loaf volume with regard to control DSC analysis.
bread made with hard WT flour. The use of a microbial transglutaminase (protein-glutamine
Blaszczack and others (2004) studied the effect of 2 α-amylases, γ -glutamyl transferase, Tgm), which catalyzes the formation of
one of fungal and the other of bacterial origin, on the texture and ε-(γ glutamyl-)-lysine crosslinks in proteins via an acyl transfer
microstructure of bread. The two α-amylases resulted in different reaction (Motoki and Seguro 1998; Larre and others 2000), has
microstructure of bread, with respect to control bread, as revealed received a great deal of interest. Tgm with or without added
by light SEM, thus staling was delayed. The authors proposed amylolytic (maltogenic bacterial α-amylase in granulate form
distinct antistaling mechanisms for the two α-amylases. [NMYL]) or non-amylolytic (PTP) had beneficial effects on hard-
ness evolution of bread obtained with white (Collar and Bollain
2005) and whole-meal flour (Collar and others 2005). Bread soft-
Xylanases (Xyns)
ness was reduced up to 16%, with respect to control bread, when
Xyns are enzymes able to retard bread staling, as reviewed by
interactive effects were tried, and the best combination was the
Butt and others (2008), to which the reader is redirected.
addition of NMYL to Tgm breads, ascribing this effect to the
A recombinantly produced Xyn B (XynB) from Thermotoga
relevant softening effect of NYML.
maritima MSB8 retarded the staling of frozen PB bread (Jiang and
Gambaro and others (2006) proposed that the addition of a
others 2008). When added to the formulation, the resulting bread
mixture of α-amylase and Xyn was able to extend the shelf-life of
had a 40% reduction in crumb firmness and retarded staling, as
brown pan bread by retarding staling, as assessed by sensory and
bread supplemented with XynB after 4 d of storage at 4 °C had the
instrumental analyses. They suggested that the mixture produces
same firmness as control bread after 1 d of storage. Data obtained
low-molecular-weight dextrins with high water retention capacity,
with DSC analysis showed that XynB was able to retard amy-
and that could be partly responsible for the lower staling rate.
lopectin crystallization. Recently, Zheng and others (2011) found
Moreover, they found a high correlation between both sensory
the right dosage to be used for 2 GH 10 Xyns, a psychrophilic
and instrumental parameters and staling rate.
(XynA from Glaciecola mesophila) and mesophilic one (EX1 from
Caballero and others (2007) studied the single and synergistic
Trichoderma pseudokoningii), with the aim to retard bread staling.
effects of some gluten-crosslinking enzymes (Tgm, glucose oxidase
Both Xyns exhibited similar antistaling effects on the bread, but
(GO), and laccase), and gluten-degrading enzymes (α-amylase,
while XynA proved to be more effective in reducing the firming
Xyn, and protease) on bread staling. They found that α-amylase,
rate, the EX1 performed better in reduction of the initial bread
Xyn, and protease were able to lower significantly the staling effect
firmness. The optimal dosage of the psychrophilic Xyn was much
promoted by Tgm and proposed different mechanisms of action
lower than that of the mesophilic counterpart, probably because
for each enzyme. In particular, they suggested that α-amylase and
the temperatures used for dough preparation and proofing were in
Xyn could have an effect on the dough polysaccharide fraction,
the range of optimum activity of psychrophilic XynA, as otherwise
while the protease may counteract Tgm-action, by a simultaneous
reported (Collins and others 2006).
action on the dough protein fraction.
Recent results of the application of a thermostable enzyme
Waters and others (2010) proposed that the highest Xyn and
cocktail from Thermoascus aurantiacus showed an antistaling effect
α-amylase activities of 5 thermozyme cocktails with different hy-
(Oliveira and others 2014). The main enzyme found on the cock-
drolytic enzyme profiles produced by Talaromyces emersonii resulted
tail was Xyn, xylose being the main product released through
in delayed staling. The enzyme cocktail B was the best in reduc-
enzyme activity after prolonged incubation, and its application at
ing crumb hardness evolution after 5 d of storage, with respect to
35 units of Xyn/100 g significantly delayed staling of bread up to
control bread.
10 d at 4 °C if compared to control loaves. On the basis of DSC
results (lower enthalpy) it was suggested that products deriving
from Xyn activity interfered with the reorganization of the amy- Others
lopectin and/or with the redistribution of water in the system, The oxidizing effect of GO was exploited for retarding staling
with a consequent retrogradation reduction. Recently Ghoshal of bread (Bonet and others 2006). When used at a concentration
and others (2013) suggested that the reduction of crystallization of 0.001%, GO delayed significantly the bread staling up to 12
and reduction of crystal growth in bread, as assessed by using n and d at 25 °C. The antistaling effect suggested was due to the large
k parameters of the Avrami equation, was caused by Xyn addition amount of total pentosans produced by GO that can associate
in whole-WT bread stored at 4 and 25 °C for 10 d, thus resulting with the glutenin macropolymer, thus leading to retention of high
in delayed staling. Measurement of thermal properties confirmed amounts of water.
the beneficial effects of Xyn, as it lowered the endothermic peak
for staling and the change of enthalpy during storage, with respect
to control bread.
Associated Mixtures of Ingredients and/or Enzymes
In this section, we will summarize the results of the main studies
dealing with ingredients and/or technological aids not included in
Enzyme mix the previous classes or combinations of different ingredients. An
The difference in mode of action of the various enzymes has interesting review on shelf-life improvement of polyols, to which
been used recently by several authors, which depended on additive the reader is redirected, has recently been published (Bhise and
or synergistic effects in order to retard staling. Kaur 2013).
Leon and others (2002) studied the effects of 2 commercial en- Wang and others (2007) reported that, when 1% of WT GHP
zyme mixtures containing α-amylase and lipase activity on staling was used, the hardness value of 3-d-old bread was equivalent to
rate. Both mixtures helped in slowing down the staling rate, espe- that of 1-d-old control bread, probably for the higher, even if
cially the blend with the higher α-amylase activity. The beneficial not significantly, specific volume and moisture content of WT
effect was attributed to a delay in amylopectin retrogradation and GHP-supplemented sample.
C 2014 Institute of Food Technologists® Vol. 13, 2014 r Comprehensive Reviews in Food Science and Food Safety 481
Bread staling review . . .
Abu-Goush and others (2008) found a beneficial effect of 11 new mathematical staling models. They found that all models
sodium-propionate in delaying staling of Arabic flat bread and cor- presented high values, the best being the rational and the quadratic,
related this result to moisture loss, starch retrogradation, and pro- thus concluding that these models are suitable to simulate staling
tein interaction effects, as revealed by near-infrared spectroscopy kinetics. The best improver formulation contained 1.27% glycerol,
data. 0.41% SSL, and 1.59% enzyme-active soy flour.
Shaikh and others (2008) tested 8 different antistaling agents on The plasticizing effect of the sorbitol on starch/gluten biopoly-
unleavened chapatti bread and measured various staling parame- mers has been described by Pourfarzad and others (2011) as the
ters such as moisture content, texture, water-soluble starch, in vitro main reason of antistaling effect of soy-fortified bread for storage
enzyme digestibility, enthalpy change, and sensory quality during times longer than 2 d and up to 5 d. The same effect was found
10 d of storage, at 4 and 29 °C. When comparing the effect of also for propylene glycol when used at 5 g/100 g flour.
the added ingredients the authors found that maltodextrin had
the highest rank at both temperatures, while the worst result was Processing Factors Affecting Staling Rate
exerted by glycerol monostearate, following the order: maltodex- Researchers have focused their attention during the last 10 y
trin > GG > α-amylase > sorbitol > XG > SSL > propylene mainly on baking technology, process parameters, and storage tem-
glycol > glycerol monostearate. Moreover, when trying 6 combi- perature, but other factors will also be reported.
nations, SSL + α-amylase gave the best texture values, suggesting Storage temperature. The effect of storage temperature on stal-
that α-amylase first breaks starch molecules, and then SSL forms ing has been reported by different authors, and the main character-
the complex with fragments derived from starch rupture. istic is a negative dependence between staling rate and temperature
The lowest amylopectin retrogradation of soy milk powder was (Colwell and others 1969).
addressed as the cause of delayed staling rate in WT–soy bread The consumer request to have “fresh” bread available at any
(Nilufer-Erdil and others 2012). This result was attributed to the time of the day (Matuda and others 2005) has stimulated the
synergistic effect of soluble fiber and partly denatured soy proteins bakery industry to exploit freezing technology and this has driven
and higher lipid content of the soy milk powder. The delay of researchers to focus their attention mainly on effects of freezing
staling was confirmed by Instron firmness measurements, although and frozen storage on bread staling, especially on dough and PB
loss moduli revealed by dynamic mechanical analysis (DMA) did samples.
not give significant differences of stiffness among formulations, A comprehensive picture up to 2008 on the effect of raw mate-
contrary to what had been reported previously by Vittadini and rial requirements, processing conditions, and baked bread quality
Volovodtz (2003). from FD and PB bread are reviewed by Rosell and Gomez (2007),
Jekle and Becker (2012) studied the effects of pH adjustment, Selomulyo and Zhou (2007), and Yi (2008), to which the reader
water, and sodium chloride addition in order to model bread tex- is redirected.
ture and staling kinetics of bread crumb. By using the Avrami Carr and others (2006) carried out a sensory comparison be-
equation and the firming rate, which gave a better square corre- tween frozen part-baked French bread (FPBFB) and fresh bread
lation coefficient, the authors managed to predict the staling rate during a week of frozen storage with daily inspections. The FPBFB
as a function of pH, NaCl, and water addition. In particular, they had a lower weight and specific volume, with respect to fresh
found an increase in the firming rate with increased NaCl concen- bread, but was rated better after 4 d of frozen storage by a con-
tration and pH reduction and a decrease when water was added sumer acceptance test (difference from control test) with respect
to the dough, probably as the change in the volume of bread had to commercial brand bread. Moreover, data on texture and sen-
a better influence on the staling rate, with respect to the effect sory analysis of FPBFB stored for a week were similar to that of
of the chemicals, since the literature well correlated the specific fresh bread. Frozen storage of PB chappati, an Indian unleavened
volume of breads with the firming rate (Axford and others 1968; flat bread, was beneficial for maintaining its quality (Gujral and
Russel 1983). others 2008). In particular, the extensibility of PB chapatti after
The addition of γ -polyglutamic acid (PGA) at 3 concentrations rebaking was very similar to that of the fresh conventionally baked
(0.5, 1.0, and 5.0 g/kg, w/w) was suggested by Shyu and others sample. The main feature was that sample of PB bread stored
(2008) to evaluate its effect on staling of WT bread. The hardness at ambient temperature or frozen (after thawing and rebaking),
value of the 6-d 1.0 kg−1 PGA stored bread was less than the value showed a significant higher extensibility when compared to the
of control bread after 1 d, thus PGA significantly reduced staling same sample of conventionally baked chapatti breads, thus giv-
rate, as also demonstrated by the decrease in cohesiveness, which ing loaves with better sensory quality than frozen conventionally
was significantly delayed by the PGA addition. baked chapatties. Yi and Kerr (2009) highlighted the influence of
Response surfaces and mathematical models were used by freezing rate (rate 1:15 °C/h, rate 2:33 °C/h, rate 3:44 °C/h, and
Gomes-Ruffi and others (2012) to show the beneficial effect of rate 4:59 °C/h), dough storage temperature (−10, −20, −30, and
the contemporary addition of SSL and of the enzyme maltogenic −35 °C) and storage duration on bread quality. They found that
α-amylase (MALTO) on both the increase of bread volume and sample frozen at the lower freezing rates and stored at the higher
the reduction of firmness, especially after 10 d of storage, when temperatures had higher specific volume, were softer, and were
the combination of 0.50 g SSL/100 g flour and 0.02 g MALTO/ lighter in color, but staled more easily, due probably to the higher
100 g flour resulted in the same firmness value as the control at damage to the starch–gluten network at slower freezing rates (Yi
day 1 of aging. The authors suggested that SSL formed complexes 2008). They noted that response of gluten structure and yeast ac-
with starch molecules, while MALTO reduced the molecular tivity to freezing rate and temperature should be balanced in order
weight of the starch molecules, thus reducing retrogradation. to find the optimal freezing conditions. Aguirre and others (2011)
Pourfarzad and Habibi-Naiaf (2012) used the positive results confirmed the existence of moisture equilibration between crumb
in changing the hardening rate of Barbari bread obtained with an and crust during bread storage, and demonstrated that storage at
antistaling liquid improver, made up of glycerol, SSL, and enzyme- −18 °C resulted in very limited water movement when compared
active soy flour, at different amounts, to test the consistency of to bread stored at 4 and 25 °C. As a consequence, water activity
482 Comprehensive Reviews in Food Science and Food Safety r Vol. 13, 2014
C 2014 Institute of Food Technologists®
Bread staling review . . .
values were almost constant in bread stored for 23 d at −18 °C. ing WT bran sourdough and an enzyme mix (α-amylase, Xyn,
They showed that the starch molecules re-associate during storage and lipase). The crumb hardness of the supplemented bread after
to give a new crystalline structure with a typical XRD B-type 6 d of storage was the same as that of white bread at day 1. The
structure and that storage at −18 °C, that is a temperature below authors used NMR, DSC, and microscopy to explain this result
the glass-transition temperature (Tg ), slowed down but did not and found fewer changes in amylopectin crystallinity and rigid-
stop the recrystallization speed, and only crystal growth occurred. ity of polymers in bran sourdough bread with enzymes, which
The effect of vacuum-cooling on the staling rate of sourdough also showed starch granules much more swollen, with respect to
whole meal flour bread was assessed by Le-Bail and others (2011). white bread, as a result of the higher water content and degrada-
Vacuum-chilled bread showed higher moisture loss, crumb hard- tion of cell wall components. In another paper, Katina and others
ness, and H of amylopectin crystals than conventionally cooled (2006b) proposed the use of surface-response methodology to op-
bread. The authors concluded that the negative effects of the quick timize sourdough process conditions aimed at improving flavor
vacuum-cooling is the result of the increased number formation and texture of WT bread. They found that combining flour with
of amylopectin crystallites and, thus, of recrystallized amylopectin. low ash content, and optimizing sourdough fermentation time,
Ronda and others (2011) studied the effect of prolonged storage staling was reduced up to 4 d. The best result was, in particular,
time on staling of PB and fully baked (FB) breads. Three parame- obtained using Saccharomyces cerevisiae sourdough fermented bread
ters, namely moisture content, firmness, and starch retrogradation for 12 h at 32 °C and with flour ash content of 0.6 g/100 g. It
as well as the Tg of the maximally freeze-concentrated state (Tg ), was also found that the fermentation time had an important linear
were considered to evaluate bread aging. The thawed and rebaked effect on softness of bread crumb. Finally, it was confirmed that
PB bread showed significantly lower amylopectin H values than higher ash content of flour increased firmness in sourdough breads
that of FB bread, and this may partially explain the similarity of fermented with Lactobacillus brevis, S. cerevisiae, or a combination
PB bread with fresh bread. The authors evidenced the need to starter (Collar and others 1994). Plessas and others (2007) pro-
select a proper frozen storage temperature, sufficiently lower than posed the use of sourdough with immobilized cells, as it resulted
Tg . Frozen storage time, moreover, resulted in a significant de- in a 3-fold delay in staling, compared to the traditional compressed
crease in firmness of PB bread crumb. Based on the obtained baker’s yeast bread. The authors hypothesized that the retention of
results, the authors proposed that hardening of bread during stor- higher moisture levels after baking and reduced moisture loss rates
age may not be related only to starch crystallization or water loss are due to the more compact texture in breads obtained with the
and developed a regression study describing how the combined suggested technique. In particular, they showed that sourdough
effect of both variables could better explain the firming evolution. breads presented lower loaf volumes for the same loaf weights, and
Majzoobi and others (2011a) hypothesized that the higher mois- fewer holes of higher size, with respect to conventional baker’s
ture content of Barbari PB flat breads after full baking was the cause yeast bread. Dal Bello and others (2007) confirmed that the higher
of delayed staling up to 72 h, with respect to control sample, and volume of bread produced by the sourdough fermentation activ-
proposed that bread crumb structure is formed completely during ity of the antifungal strain Lactobacillus plantarum FST 1.7 and of
the part-baking stage, while staling occurs in PB bread during Lactobacillus sanfranciscensis LTH 2581, with respect to chemically
storage at ambient temperature, even if full-baking leads to the or nonchemically acidified bread, delayed crumb staling up to 3 d.
disappearance of many signs of staling, thus the resulting bread has Additionally, the L. plantarum FST 1.7 revealed inhibitory activity
softer texture. Finally, they suggest storing the part-baked bread at against Fusaria.
frozen temperature for no more than 2 mo to reduce deterioration Fadda and others (2010) found that durum WT bread produced
of bread caused by the growth of ice crystals. In a subsequent pa- with sourdough at a dose higher than 10% significantly lowered
per Majzoobi and others (2012) recommend the addition of 15% and slowed crumb-firming kinetics, as assessed by TA and DSC
WT germ for the general sensory improvement of Barbari bread, results, the latter used with the Avrami equation, provided gluten
although that did not manage to retard staling. and yeast were added.
In 2 separate papers Karaoglu (2006) and Karaoglu and Recently, Tamani and others (2013) associated the increased
Kotancilar (2006) evidenced the influence of par-baking on EPS production during dough formation following the inocu-
quality of WT bran and white breads, respectively, supplemented lation of ropy LAB starter cultures (Lactobacillus delbrueckii subsp.
or not with calcium propionate, during chilling storage (4 °C) bulgaricus LB18; L. delbrueckii subsp. bulgaricus CNRZ 737, and L.
up to 21 d. Both papers gave similar results, which were a softer delbrueckii subsp. bulgaricus 2483) with increased bread volume and
bread crumb, with respect to a control group, in breads PB for 10 reduced staling over 5 d of storage, with respect to the control
min, rebaked, and stored for 7 and 14 d. bread, while 1 nonropy LAB (Lactobacillus helveticus LH30) did not
result in beneficial effects. The authors suggested that the higher
Sourdough fermentation levels of EPS obtained with LAB may have resulted in greater
Sourdough fermentation has been known since ancient times water retention, leading to the softer crumb structure of these
and, among the beneficial effects, reduction in staling has been breads, even if they evidenced that the EPS production did not
reported and recently discussed in 2 reviews (Arendt and others correlate with the extension of shelf-life, thus their effect was more
2007; Chavan and Chavan 2011), to which the reader is redirected. qualitative than quantitative.
The different metabolites produced by lactic acid bacteria (LAB)
have proved to have a beneficial effect on texture and staling. EPS, Baking and fermentation
for example, are a valid and economic alternative to hydrocolloids, It has been reported that both baking time and temperature
while organic acids affect the protein and starch fractions and affect the quality and staling rate of bread (Seetharaman and oth-
reduce the pH that results in an increase in protease and amylase ers 2002). Patel and others (2005) studied the effects of the use
activities of the flour, thus reducing staling. of different ovens and dough size, when baking at constant tem-
Katina and others (2006a) managed to delay bread staling at 3 perature for varying times, on texture, thermal properties, and
and 6 d of storage, with respect to white WT bread, by combin- pasting characteristics of products. Breads baked at the lower
C 2014 Institute of Food Technologists® Vol. 13, 2014 r Comprehensive Reviews in Food Science and Food Safety 483
Bread staling review . . .
heating rates had lower amylopectin recrystallization, rate of bread adjust the firmness parameters to simple curvilinear equations and
firmness, and amount of soluble amylose. Similar results were ob- obtained high correlation coefficients (>90%). Ozkoc and others
tained by Mouneim and others (2012). Baking temperature and (2009) compared different baking methods, namely conventional,
time affected some physical properties of bread from composite microwave, and infrared-microwave combination, in order to as-
flour made by mixing cassava and WT flour at a ratio of 10:90 sess staling kinetics of hydrocolloid-supplemented breads during
(w/w) as revealed by central composite rotatable experimental de- 120 h of storage, by using several methods, namely TA, DSC, RVA,
sign (Shittu and others 2007). Both the baking temperature and X-ray, and Fourier transform infrared spectroscopy (FTIR). The
time, among others, influenced the dried crumb hardness, due to starch retrogradation of breads obtained with a combination oven
the complex effect of temperature and time combination, but the was similar to that of conventionally baked ones, as revealed by
developed 2nd-order response surface regression equations could H values and FTIR outputs, thus leading the authors to postu-
not predict satisfactorily most of the measured properties, thus the late that it was possible to produce breads by combination heating
authors proposed further studies to optimize the cassava and WT with a staling rate similar to that of conventionally baked ones.
flour bread baking process. Three different heating temperatures Moreover, data from RVA and X-ray showed that the rapid staling
corresponding to 3 heating rates were also tested by Le-Bail and rate typical of microwave baking can be mitigated by infrared-
others (2009) with an innovative protocol in which a degassed microwave combination heating. As expected, the addition of a
piece of dough was baked in a miniaturized oven, in order to XG–GG blend to the formulation retarded staling.
compare it with traditional dough. Hardening of the crumb oc-
curred after retrogradation of amylopectin, as revealed by calori- High-hydrostatic-pressure processing (HPP)
metric tests, and higher baking kinetics resulted in faster staling This unit operation may change structural and functional prop-
rates. Additionally, the relative Young modulus, expressed as the erties of proteins and cereal starches and is being investigated to
ratio of the modulus of the cellular crumb compared with the improve quality of breads made with flours alternative to WT.
modulus of the degassed crumb, was proportional to the square of In a fundamental study on the use of HPP to improve the
the relative density of the crumb. In a further paper Le-Bail and bread making performance of oat flour, Huttner and others (2010)
others (2012), working with a degassed sourdough, confirmed the subjected oat batters to 3 levels of HPP (220, 350, and 500 MPa)
previously obtained results and gave more explanation on the ef- and the treated samples replaced untreated oat flour in an oat bread
fect of prolonged baking on staling rate, that was an increase of recipe, by 10%, 20%, or 40%. Staling rate, as assessed by a TPA
the amount of amylose leaching from the starch granule, lead- crumb hardness test, was reduced when 10% to 40% oat batter
ing to a higher Young’s modulus of the crumb at the end of treated at 200 MPa was used, if compared to the control. The HPP
staling. treatment at 200 MPa weakened the proteins, affected the moisture
Different heating rates were recently associated with water va- distribution, and also influenced the interactions between proteins
por permeability (WVP), effective moisture diffusivity (Deff ), and and starch, which caused a decrease in the staling rate of the oat
sorption of bread crust and crumb (Besbes and others 2013). The bread. Opposite results were presented in another paper published
authors showed that baking at 240 °C gave both crust and crumb some months later (Vallons and others 2010). The authors replaced
with higher moisture diffusivity coefficient and that the crust had 2% or 10% of a sorghum bread recipe with sorghum batters HPP-
a higher WVP than that of sample baked at 220 °C. They pro- treated at 200 and 600 MP and found that breads containing 2%
posed a more pronounced porosity of crumb and crusts of breads sorghum treated at 600 MPa had slower staling rates than control.
baked at the higher temperature, as revealed by porosity values and More recently, Angioloni and Collar (2012) worked with fixed
scanning electron microscopy (SEM) determinations, as the cause amounts of oat, millet, and sorghum HPP-treated flours (350
of the obtained result. Purhagen and others (2012) concluded that MPa), which replaced (60% for oat, 40% for the other 2) WT
breads obtained with different fibers (fine durum, oat bran, rye flour. Half of the control bread was prepared by applying HPP to
bran, and WT bran) baked in pan remained softer after 7 d of 50% of WT flour. Results indicated that HPP-treated WT and
storage, with respect to free-standing baked sample, and attributed oat breads lowered final values of crumb hardness and Avrami
this to the lower specific volume of pan-baked breads due to their exponent, thus giving softer breads with slower staling kinetics,
high water content. Moreover, pan-baked loaves lost less water with respect to control bread.
during storage, with respect to free-standing sample, probably be-
cause of the smaller crust area of these loaves. The difference in Measurement Methods
staling behavior between the 2 baking methods was not attributed, The results reviewed above refer to one or more measurement
however, to starch retrogradation, while the influence of fibers was methods to assess bread staling, but there has not been up to now
small, if compared to the baking method, thus confirming data ob- a methodology that allows a complete measurement of the staling
tained in another paper in which other antistaling agents, namely phenomenon to the same extent as that described by a consumer
α-amilase, distilled monoglyceride, and lipase, were compared to (Sidhu and others 1996). Different specific reviews before that of
the baking method (Purhagen and others 2011a). Gray and Bemiller (2003) have dealt with the methods used to
The effect of fermentation on the firming kinetics could not be assess the rate and/or degree of staling such as those mentioned
explained only by its effect on volume, but also with the presence by Maga (1975), Kulp and Ponte (1981), and Ponte and Ovadia
of different enzymes, such as amylases, proteases, or lipases that, (1996). In most cases bread staling, apart from the more simple
alone or in combination with other enzymes, may help in reducing and direct TA, is indirectly measured as the extent of starch ret-
the firming rate in white or wholemeal bread, thus longer fermen- rogradation, as also reviewed by Karim and others (2000). An
tation times enhanced the action of the enzymes, with a resulting interesting review, moreover, revisited crumb texture evaluation
reduction of the staling rate (Gomez and others 2008). The higher methods (Liu and Scanlon 2004), while another one summarized
the yeast dose, the higher the quantity of dough enzymes previ- the more frequently used analytical methodologies for assessing
ously cited. Temperature of fermentation, on the other hand, had a bread staling (Choi and others 2010). In the following sections
minor impact on bread staling. Moreover, the authors managed to the major reports dealing with new methodologies and/or new
484 Comprehensive Reviews in Food Science and Food Safety r Vol. 13, 2014
C 2014 Institute of Food Technologists®
Bread staling review . . .
applications used to measure bread staling during the last 10 y will it was able to study accurately amylopectin retrogradation and to
be reviewed. obtain a very good correlation with DSC data when looking for
protein and temperature effects on amylopectin retrogradation de-
Thermal analysis velopment, even if it showed difficulty in measuring the changes of
Bollain and others (2005) proposed small dynamic defor- the amylase–lipid complex during storage. The authors proposed
mation and large static deformation methods to evaluate the 550, 970, 1155, 1395, and 1465 nm as important wavelengths of
thermodynamic and physical–mechanical changes of enzyme- NIRS and concluded that amylopectin retrogradation was prob-
supplemented white or whole bread during staling. They suc- ably the main factor in bread staling and that the amylase–lipid
cessfully detected rheological changes of bread, as influenced by complex contributed little to bread staling after 1 d of storage.
recipe and storage time, with dynamic thermomechanical analysis Cocchi and others (2005) coupled middle-infrared spectroscopy
(DTMA) in the compression mode. They detected that the onset (MIR) with principal component analysis (PCA) to follow bread
frequency (f0 ) and the rubbery or plateau moduli (E ) rose as the shelf-life in a rapid and affordable way. Spectra of breads stored up
bread aged in a similar way to the hardening and firming curves. to 7 d at ambient temperature were acquired in attenuated total
Moreover, relationships between the dynamic (DTMA) and static reflection mode with an FTIR spectrometer, normalized and then
(TA) methods were found. subjected to PCA. The authors revealed that the 1st PC increased
Ribotta and Le Bail (2007) used DSC and DMA to study bread with aging of samples and that the more influential variables on
staling. DSC evidenced water migration from the crumb to the PC1 corresponded to spectral regions attributed to typical starch
crust and changes of water properties as initial and onset temper- bond vibrations.
ature of ice melting decreased significantly after 1 d and freezable Pikus and others (2006) proposed for the first time the small-
water (FW) and unfreezable water (UFW) decreased and increased, angle X-ray scattering (SAXS) method to study bread staling. The
respectively, as a consequence of aging. DSC results suggested the authors, by using fresh dry and fresh water suspension samples,
existence of a possible 2nd transition, due to ice-melting transition found that bread staling is accompanied by significant electron
being diverted to lower temperatures. The authors proposed that a density changes, indicating that there were significant changes at
concomitant water migration from the crumb to the crust and an the nanoscale level during the staling process. They suggested,
incorporation of water molecules into the starch crystalline struc- by analyzing results obtained with the dynamics of the scattering
ture, developing after bread staling, may account for the decrease intensity changes in the bread samples, along with those of SAXS
in FW after 4 d of storage at 4 °C. Moreover, they suggested investigations on native starch, that SAXS scattering changes for
that some water molecules were incorporated in the crystalline the dry samples originated mainly from the gluten phase, while
lattice when starch crystallized. DMA analysis showed significant for water suspension samples they were mainly from the starch
changes in the thermo–mechanical profile of the crumb during matrix. The authors concluded that a comparison of results of
staling, as aged breads contracted at a lower rate during cool- SAXS with data obtained with other methods, on the same bread
ing, but they evidenced a greater deformation during freezing and sample, would be interesting.
higher retraction within the complete cooling–freezing cycle, thus Piccinini and others (2012) proposed, for the first time the
suggesting that the higher matrix rigidity, a consequence of the use of NIR Fourier-transform-Raman spectroscopy to monitor
higher amount of retrogradated starch, affected contraction capac- starch retrogradation in stored hard-WT bread and, with the help
ity. The authors postulated that interactions during the hydration of TA data, to follow bread staling for 20 d. The authors found,
of the gluten network might explain the latter phenomenon. by applying the 2D correlation analysis applied to the Raman
spectra of bread crumb during storage, that both the peak shift
Infrared spectroscopy and narrowing of the band at 480 cm−1 during retrogradation
Near-infrared reflectance spectroscopy (NIRS) was used to ob- correlated well with the crumb-firming data obtained using the
tain spectra during staling of bread and the results were compared stress relaxation tests and that during starch retrogradation a new
with those obtained by TA (Xie and others 2003). Results showed band peaking at 765 cm−1 appeared.
that NIRS spectra were highly correlated with firmness values
assessed with the more common TA. Moreover, the authors ev- NMR spectroscopy
idenced that NIRS measurements had a better correlation with Curti and others (2011) used 1 H NMR relaxometry and, for
storage time and also lower batch variability, with respect to TA- the first time in bread, the 1 H NMR fast field cycling (FFC) tech-
derived data, thus NIRS was suggested as a better tool than TA to nique to follow the changes in 1 H T1 relaxation in the 0.01 to 20
study bread aging, probably because NIRS may follow both phys- MHz frequency range, in order to check for the interactions of
ical and chemical changes occurring during the staling process, water molecules with paramagnetic and large-sized macromolec-
while TA was limited to the only aspect of firmness evolution. In ular system during bread staling. 1 H T1 relaxation data at 20 MHz
a further paper, Xie and others (2004) proposed the use of NIRS confirmed previous results, while studies conducted at a lower fre-
as a fundamental tool to study bread staling with the help of DSC, quency (0.52 MHz) evidenced, for the first time, the presence of
as well as the effects of starch, protein, and temperature (stor- two T1 proton populations, which were tentatively attributed to
age at 12.5 or 31.5 °C) on bread staling. DSC data showed that protons of the gluten domain at early storage times. The authors
temperature strongly affected the staling rate, while the protein suggested that the use of the 1 H NMR FFC technique at different
contribution was limited, if compared to temperature during 4 d frequencies may be an additional way for monitoring molecular
of storage. Using the enthalpy ratio between bread supplemented dynamics in bread and therefore a new valuable instrument to help
with starch and sample produced with starch–protein, it was pos- understand the bread staling phenomenon.
sible to conclude that protein might retard bread staling not only Bosmans and others (2013) used H NMR relaxometry, along
by diluting starch (Kim and D’Appolonia 1977; Every and others with DSC and wide-angle XRD, to better elucidate the relation-
1998), but also by interfering with amylopectin retrogradation. ship between biopolymer interactions, water dynamics, and crumb
NIRS was found to be very useful in studying bread staling, as texture evolution during 168 h of storage of bread. The NMR
C 2014 Institute of Food Technologists® Vol. 13, 2014 r Comprehensive Reviews in Food Science and Food Safety 485
Bread staling review . . .
analysis allowed finding 6 proton populations in bread crumb and ships between mechanical behavior of pan bread, supplemented
from the NMR profiles of bread crumb they were able to de- or not with an amylolytic enzyme, a nonamylolytic enzyme, and
duce the extent of formation of both amylopectin crystals and a combination of the 2, and loss of sensory quality during 20 d in
of crumb firmness. On the basis of data obtained, they con- storage. TPA at 40% and 80% was proposed for the first time as
cluded that the increase in crumb firmness of stored bread was well as a new penetration test. The authors positively correlated
caused by a combination of different events that were amylopectin hardness with sensory “difficulty in swallowing,” “crumbliness,”
retrogradation and the formation of a continuous, rigid, crystalline “hardness,” and “oral dryness,” and negatively correlated it with
starch network that included water in its structure. They also no- sensory “cohesiveness,” “softness,” and “size of soft zone,” while
ticed moisture migration from gluten to starch and from crumb to these parameters correlated well also for springiness and cohe-
crust, resulting in additional reduction of moisture in the gluten siveness detected at 80% TPA, thus evidencing that TPA values
network, with the consequence that the subsequent increase in obtained at the compressions resulted in greater sample distortion
stiffness contributed to the increase in crumb firmness. and gave information that was better correlated to sensory per-
ception. Finally, the analysis of the penetration profiles gave data
X-ray crystallography that were very useful to complement the TPA results, in order to
Del Nobile and others (2003) developed a mathematical model assess bread staling.
able to predict the starch retrogradation kinetics of durum WT Angioloni and Collar (2009b) suggested the complementar-
bread in order to link it to the crumb staling. Two equations were ities of instrumental static (TPA, firmness, and relaxation test)
proposed dealing with data obtained with wide-angle XRD (starch and dynamic (innovative oscillatory test) analyses with empirical
retrogradation) and compression tests (crumb firming process), and sensory characteristics in assessing commercial whole and white
related to samples held at 5 °C and 2 water activity values, in order bread quality during a 10-d storage period, although the 2 differ-
to accelerate the test. The proposed model fitted well with the ent approaches investigated the bread characteristics at molecular
obtained results; moreover, the authors evidenced that lowering or macroscopic level. In particular, the authors found that static re-
the water activity value resulted in a higher overall starch crystal laxation parameters initial force (F0 ), momentary force at time (t)
growth rate, due to the increase of the starch nucleation rate. F(t), constants related to stress decay k1(s) rate and residual stress at
X-ray patterns were studied with different methods, namely rel- the end of the experiment (k2t), and dynamic (stress) bread crumb
ative crystallinity, total mass crystallinity grade (TC), B-type mass rheological attributes were correlated well, thus both techniques
crystallinity grade, and V-type mass crystallinity grade, in order to were useful in evaluating crumb textural characteristics of fresh
increase knowledge of the relationship between starch crystallinity and staled breads. Moreover, the sensory attributes (softness) and
and bread staling during 7 d of storage at 4 °C (Ribotta and oth- the overall acceptability were negatively correlated with either dy-
ers 2004b). The authors pointed out that: (a) fresh baked bread namic stress or static F0 . The authors concluded that the obtained
contained only a V-type structure, while the B-type structure ap- results were quite promising for a proper bread crumb quality as-
peared after 24 h and increased during bread staling; (b) TC and sessment, as the novel proposed approaches gave data with better
relative crystallinity significantly increased during the first 24 h, accordance with consumer awareness.
then slightly decreased, thus indicating the appearance of the B-
type structure; (c) TC and relative crystallinity decreased at the end Electrical impedance
of aging, which is associated with an increased degree of ordering Bhatt and Nagaraju (2009) developed an instrument working
of the amorphous phase caused by staling. They suggested that with electrical impedance to assess the electrical properties of
staled bread showed reformation of the double helical structures WT bread crumb and crust, and they investigated changes in
of amylopectin and a reorganization, during aging, into crystalline electrical impedance behavior during 120 h of storage with the
regions that imparted rigidity. With this in mind, they concluded use of multichannel ring electrodes. Variations in crust capacitance
that amylopectin retrogradation is an essential step to consider to showed that there was a sharp increase in value after 96 h of storage
better understand bread staling. at 17.6% moisture content, so after that period a glass transition
Colorimetry occurred with a content of more than 17.6% of moisture at room
Popov-Raljić and others (2009) used, for the first time, an temperature. On the other hand, the resistance measurements of
MOM-color 100 tristimulus photo colorimeter, in CIE, CIELab, crumb showed a decrease during staling, thus revealing that the
ANLAB, and Hunter systems to correlate crust color changes and starch crumb recovered its crystallinity during the storage time
staling of bread of different compositions packed in polyethylene of 120 h. Data on crust capacitance and crumb resistance were
film during 3 d at 20 °C. The color of 3-d-stored bread sam- validated by results obtained with DSC analysis (variation in glass
ples was always lighter, as the stored breads showed higher average transition temperature and enthalpy). The authors concluded that
reflectance, with respect to just baked loaves. The authors hy- the proposed instrument was suitable for rapid and nondestructive
pothesized the moisture loss as the cause of this color change and, measurement of electrical properties of bread at different zones
by fitting the values of average reflectance with a curve describ- with minimum error, thus enabling to study staling at crust and
ing the dependence of average reflectance with storage time, they crumb simultaneously.
found a correlation coefficient of 0.99, thus they concluded that
the change in color is the direct consequence of staling (Note: it Mixed instrumentation
would be more useful to correlate crust color changes with objec- Primo-Martı́n and others (2007) gave new insight on staling of
tive bread staling measurements, such as hardness, more than with bread crust by using a wide range of measurement techniques,
time). namely, confocal scanning laser microscopy, wide-angle X-ray
powder diffraction, polarized light microscopy, solid-state 13C
Rheological methods cross-polarization-magic-angle spinning NMR, and DSC. The
Textural assessment of staling has been reviewed by Chung and authors found that baking resulted in gelatinization of only 60%
others (2003). Fiszman and others (2005) investigated the relation- of the crust starch, and this fraction retook its crystallinity after a
486 Comprehensive Reviews in Food Science and Food Safety r Vol. 13, 2014
C 2014 Institute of Food Technologists®
Bread staling review . . .
long time, compared to crumb. The authors, thus, concluded that hydrostatic pressure, ultrasound processing, pulse-light technol-
staling of the crust cannot be ascribed to amylopectin retrograda- ogy, and others), and their effects on the retardation of bread
tion that was measurable only after 2 d of storage, while loss of staling.
bread crust freshness happened before 1 d of storage, as already
reported by Primo-Martı́n and others (2006). Acknowledgments
A very interesting application was that proposed by Botre and The authors acknowledge the financial support of follow-
Garphure (2006) who used a tin oxide sensor array and self- ing European Institutions: Regione Autonoma della Sardegna,
organized map (SOM)-based E-nose for analysis of volatile bread Legge 7, project title “Ottimizzazione della formulazione e
aroma, in order to correlate the obtained data with bread freshness della tecnologia di processo per la produzione di prodotti da
and, thus, predict staling. Data obtained on bread stored for 5 d forno gluten-free fermentati e non fermentati,” and Consejo
at 25 °C over 3 wk and purchased by 3 producers showed that Superior de Investigaciones Cientı́-ficas (CSIC), and Ministerio
the E-nose was able to predict freshness or staleness of bread with de Economı́-a y Competitividad (Project AGL 2011-22669) of
an accuracy of up to 97%, when using data sets and the SOM Spain.
network of the same week, while this value dropped from 75% to
85% when considering the 3 wk. Moreover, when different bread
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