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International Journal of Biological Macromolecules

27 (2000) 1 – 12
www.elsevier.com/locate/ijbiomac

Review
Annealing of starch — a review
Richard F. Tester *, Stéphane J.J. Debon
Food Research Laboratories, School of Biological and Biomedical Sciences, Glasgow Caledonian Uni6ersity, Glasgow G4 0BA, UK

Received 22 July 1999; accepted 1 December 1999

Abstract

Annealing processes, involving specific heating protocols, have been used by man for centuries to impart desirable properties
to materials — especially metals and particularly tools and weapons. The terminology has also been applied to biopolymers such
as starches, where the effects of the processing have been known for decades although the molecular basis has not been at all well
understood. Because of the marked effect the annealing process has on starch functionality and consequently industrial
applications, it is critical that the underlying molecular events are understood. This review is an attempt to clarify the process of
starch annealing with an emphasis on data generated in the authors’ laboratory. © 2000 Elsevier Science B.V. All rights reserved.

Keywords: Annealing process; Heat–moisture treatment; Starch

1. Background and definitions Corke [4] have helped to clarify the situation. They
state that annealing represents ‘physical modification of
The annealing process, when related to starches, has starch slurries in water at temperatures below gelatini-
been variously described. Both annealing and heat– sation’ whereas heat–moisture treatment ‘refers to the
moisture treatments are related processes, where the exposure of starch to higher temperatures at very re-
starch to moisture ratio, temperature and heating time stricted moisture content (18–27%)’.
are critical parameters to control. Jacobs and Delcour These authors propose that the terminology is stan-
[1] have discussed the difference between annealing and dardised — which has implications in terms of the
heat –moisture treatment of starch. They state that definition of gelatinisation. Hence, the following defini-
treatments in excess (\60% w/w) or at intermediate tions are proposed with respect to starch (and related
(40 – 55% w/w) water contents represent annealing while polymeric systems).
treatments below 35% (w/w) can be described as heat–
moisture treatment. Also, they state that both processes 1.1. Glass transition temperatures
occur at above the glass transition temperature (Tg) but
below the gelatinisation temperature. However, the Glass transition temperatures are very important
term heat–moisture is often used to describe high tem- parameters that affect polymeric physical properties.
perature treatments, like 100°C (up to 16 h at 27% The transition is similar to a second-order thermody-
moisture) [2]. Stute [3] has also discussed the difference namic transition and has been well described by Bili-
between annealing and heat – moisture treatments, ac- aderis et al. [5]. The term describes the temperature
knowledging that for work conducted in the early part induced transition of an amorphous glassy polymer
of last Century, annealing and heat – moisture were system to a progressively more rubbery state when it is
used as synonymous terms. More recently Collado and heated (usually in the presence of a solvent/plasticiser,
when applied to polysaccharides). In the case of com-
* Corresponding author. Tel.: +44-141-3318514; fax: + 44-141-
pletely glassy polymers, Tg is relatively distinct, where
3313208. an inflection (increase) in the specific volume and en-
E-mail address: r.f.tester@gcal.ac.uk (R.F. Tester) thalpy as a function of temperature occurs and is

0141-8130/00/$ - see front matter © 2000 Elsevier Science B.V. All rights reserved.
PII: S 0 1 4 1 - 8 1 3 0 ( 9 9 ) 0 0 1 2 1 - X
2 R.F. Tester, S.J.J. Debon / International Journal of Biological Macromolecules 27 (2000) 1–12

reflected in discontinuity in the specific heat capacity 1.2. Annealing


(Cp). Because starch contains both amorphous and
crystalline material, the exact thermal event represent- Annealing represents the physical reorganisation of
ing Tg is difficult to detect. However, high sensitivity starch granules (or appropriate polysaccharide matrices
differential scanning calorimetry (DSC) has allowed for like amylose–lipid complexes) when heated in water (or
the measurement of Tg in amorphous and native appropriate plasticiser) at a temperature between Tg
starches with various levels of crystallinity [6 – 8]. Water and the onset of gelatinisation (To) of the native starch
is a very effective plasticiser of amorphous starch (and (or polymeric system). It is recognised that annealing
hence Tg), where the ratio of starch to water is critical can be associated with partial gelatinisation. However,
with respect to the temperature at which Tg occurs (Fig. these authors believe the definition should be applied
1). only where gelatinisation does not occur and hence To

Fig. 1. State diagram of the starch–water system. The experimental data for the glass transition (Tg) are from amorphous starch [8] while the
theoretical Tg is derived from the Couchman–Karasz equation [9]. The experimental data for the melting transition (Tm) are from DSC
(Tconclusion) of wheat starches at different moisture content [10 – 13]. The theoretical Tm is fitted from the Flory – Huggins equation [14].
R.F. Tester, S.J.J. Debon / International Journal of Biological Macromolecules 27 (2000) 1–12 3

Fig. 2. DSC thermograms of a commercial wheat starch (BDH 30265): (a) native; (b) after annealing in excess water (45°C, 100 days).

must not be exceeded. In addition, according to this an irreversible molecular transition. This irreversible
definition, the enthalpy of gelatinisation post-annealing step involves dissociation of double helices (most of
cannot be less than for the native starch. Annealing which are in crystalline regions) and expansion of gran-
leads to elevation of starch gelatinisation temperatures ules as the polymers (and granule interstices) hydrate.
and sharpening of the gelatinisation range (defined The onset temperature (Tonset or To, typically  45°C)
below) as shown in Fig. 2. by DSC reflects the initiation of this process, which is
The annealing process has important industrial impli- followed by a peak (Tpeak or Tp, typically 60°C) and
cations. Starches may be deliberately annealed to im- conclusion (Tconclusion or Tc, typically 75°C) temperature
part novel processing characteristics. However, there (Fig. 2). After Tc, all amylopectin double helices have
are few commercial processes where annealing may be dissociated, although swollen granule structures will be
justified in terms of energy and time to generate retained until more extensive temperature and shear
starches with higher gelatinisation temperatures — es- have been applied. Beyond  95°C an amorphous gel is
pecially when many inexpensive chemical processes can
formed. The temperature range Tc –To represents the
be employed, over a short time frame, to selectively
gelatinisation period.
modify starch characteristics. Often annealing is
After gelatinisation, a-glucan chains re-form double
achieved unintentionally. One example is the wet
helices if the conditions are desirable. This process —
milling of maize when used to extract starch.
retrogradation — occurs when, for example, bread
1.3. Gelatinisation stales. Sometimes, annealing type processes are con-
fused with retrogradation. However, annealing of
Gelatinisation is a term used to describe the molecu- starch granules is a process that retains granular struc-
lar events associated with heating starch in water. ture and original order. Retrogradation occurs as
Starch is converted from a semi-crystalline, relatively amorphous a-glucan chains form double helices and,
indigestible form to (eventually) an amorphous (readily perhaps eventually, align themselves in crystallites.
digestible) form. The gelatinisation process (in excess Blanshard has discussed gelatinisation type processes
water) is believed by these authors to involve primary in depth elsewhere [15–17], and how interactions be-
hydration of amorphous regions around and above Tg, tween starch and other groups (especially water and
with an associated glassy-rubbery transition. This in solutes) modify the temperature driven transitions.
turn facilitates molecular mobility in the amorphous Readers are referred to these publications for more
regions (with reversible swelling) which then provokes detail.
4 R.F. Tester, S.J.J. Debon / International Journal of Biological Macromolecules 27 (2000) 1–12

1.4. Heat–moisture custard). Drier food products have often been processed
under high moisture conditions and equally, little granule
Heat–moisture treatments represent the control of form is apparent (e.g. wafer biscuits). Where water is
molecular mobility at high temperatures by limiting the limiting, however, like fat rich shortbread biscuits, essen-
amount of water and hence gelatinisation. In common tially native granule form is apparent under the micro-
with annealing, physical reorganisation is manifested. scope-although this starch has presumably been heat
The low levels of water in the system lead to an elevation moisture treated.
of Tg — the trigger for polymeric reorganisation — as The Tg must be reached or exceeded for annealing to
discussed below. Hence, high temperatures are required occur. Many authors accept that this is a prerequisite of
to cause physical reorganisation within granules. the annealing process [17,22,23]. Indeed, the annealing
Heat–moisture treatments of starches may be con- process has been discussed in terms of the process itself
ducted deliberately by industry to impart novel charac- improving Tg without facilitating the gelatinisation pro-
teristics. One example is the pre-treatment of starches for cess [17]. If starches are heated at progressively higher
infant foods. Other examples of industrial applications temperature above Tg, they do eventually completely
of the process include processing of potato starch to gelatinise, having gone through an early phase involving
replace maize starch in times of shortage, creation of enhanced mobility of amorphous regions. It is logical
excellent freeze–thaw stability and improvement of the that this phase is comparable to the phase that initiates
baking quality of potato starch [4]. and forms part of the annealing process [19,24–27].
Perhaps because of the difficulty associated with mea-
suring Tg, some authors claim that starches can be
2. Relationship between Tg on both annealing and annealed below Tg [28,29]. However, this would mean
gelatinisation that structural reorganisations of the crystalline compo-
nent of starch granules occur independently of reorgan-
We view the gelatinisation process as a co-operative isation of the amorphous phase. This is an almost
event between amorphous and crystalline regions in impossible situation to imagine in view of the relatively
starches. In unlimited water, amorphous regions imbibe impenetrable nature of these regions by water molecules,
water as the starch granules are progressively heated. with no associated passage through (and associated
Perhaps the relative large amorphous growth ring type reorganisation of) amorphous regions.
regions are the primary amorphous regions to hydrate, It is relatively straightforward to measure the gelatini-
followed by amorphous lamellae ‘sandwiched’ between sation endotherm of starch using DSC, although this is
the crystalline lamellae. The plasticisation of the amor- not true of Tg. Whilst Tg has reportedly been determined
phous lamellae and annealing of double helices is repre- prior to gelatinisation [24], it is in fact very hard to detect
sented in Fig. 3. and quantify [5,22] — unless high sensitivity DSC is used
The absorption of water into amorphous regions is at low moisture contents [6–8]. Primarily this is because
certainly possible, as for example, potato starch can it is both small and submerged into the thermogram
reversibly absorb up to 0.53 g water/g dry starch before baseline. Model polymeric systems, for example polyan-
the irreversible steps within the gelatinisation process are hydroglucose compounds [30] have, however, been useful
exceeded [17]. The water induces a transition of the in developing an understanding of how a-glucan struc-
amorphous regions from a rigid glassy state to a mobile ture itself moderates Tg. For example branched regions
rubbery state which in turn facilitates the hydration and (which are also found in the amorphous zones of starch)
dissociation of double helices in crystallites. The dissoci- seem to depress Tg similarly to plasticisation by small
ation of the crystallites begins after Tg of amorphous molecules [30].
regions, and at this temperature (To), limited dissociation
of amylopectin double helices (most of which are in
crystallites) is associated with limited swelling of gran- 3. Effects of starch-to-moisture ratio on Tg and
ules. Gelatinisation proceeds as the temperature is in- annealing
creased, progressively uncoiling all the double helices and
converting crystalline material to amorphous material. The original work of Gough and Pybus [31], which was
If water is sufficiently low so as to restrict gelatinisa- the first to describe how wheat starch could be annealed
tion, the primary gelatinisation endotherm (G) develops by heating in excess water at 50°C, showed that gelatini-
a high temperature trailing shoulder (M) [19,20], as sation temperatures could be increased while the
shown in Fig. 4. As the volume fraction of water (61) is gelatinisation range could be sharpened. Many studies
reduced to B 0.45, the shoulder becomes distinct and have been conducted on the effects of annealing
represents the only endotherm observed. In high mois- on different starches, with different starch-to-moisture
ture food systems, starch granules are completely gela- ratios and different storage times [23,29,32–46]. Some-
tinised and often no granule form is discernible (e.g. times, the annealing process is conducted as a
R.F. Tester, S.J.J. Debon / International Journal of Biological Macromolecules 27 (2000) 1–12 5

Fig. 3. Pictorial representation of the effect of hydration and subsequent annealing on the semi-crystalline lamellae (amylopectin double helices
are represented as rectangles): (a) dry starch with glassy amorphous regions; (b) hydrated annealed starch with rubbery amorphous regions
(adapted from [18]).
6 R.F. Tester, S.J.J. Debon / International Journal of Biological Macromolecules 27 (2000) 1–12

Fig. 4. Influence of water content on the differential scanning calorimetry thermograms profile of potato starch: (a) the onset ( ) and conclusion
() gelatinisation temperatures (adapted from [21]); (b) the corresponding DSC thermograms (adapted from [19]).

single event (single-step) whilst at other times it is ceeded 60%. Although annealing could be initiated at
conducted as two starch – water/temperature/time events 15°C below To by DSC, the effect was more marked the
(double-step) or even many individual steps (multi-step) closer the annealing temperature was set to (below) To.
as discussed elsewhere [1]. This double or multi-step Similar studies have been conducted on starches of
approach is often used to promote annealing without different botanical origins [32,40] and demonstrate the
gelatinisation, and the double step process potentially additional complication of species specific variation.
produces higher gelatinisation temperatures than the During annealing of starches, there are in essence two
single step process [1]. The lack of standardisation of thermally driven processes which are intimately related
annealing conditions makes it difficult to compare re- and reflect the moisture content of the system — the
sults between the different studies. elevation of Tg and gelatinisation temperatures (espe-
The effect of the starch-to-water ratio, temperature cially To). Low moisture causes elevation of (the rela-
and time on annealing of wheat starch has been investi- tively unplasticised) Tg of starches [6,7,22,47] and model
gated in detail [23]. This study demonstrated how criti- polymeric systems [30,48,49] which, in the case of
cal the interrelationship of these parameters is. The starch, intimately reflects the increase in gelatinisation
annealing process could be initiated when the moisture temperatures. Indeed, the elevation of Tg implies a more
content exceeded 20% by weight, (because Tg is around glassy state and hence reorganisation of amorphous
room temperature when this moisture content is ex- regions. This is associated with improved order of
ceeded, Fig. 1) but was restricted (in terms of its effect crystalline regions (below). The situation with respect to
on increasing gelatinisation temperatures) unless it ex- Tg of starch in food systems is very complex because
R.F. Tester, S.J.J. Debon / International Journal of Biological Macromolecules 27 (2000) 1–12 7

Fig. 4. (Continued)

of the raft of potential interactions. More details con- of starches (which can also be modelled using potato
cerning (general) glass transitions in model systems can microtubers [53]), and parallels have been drawn be-
be found elsewhere [50]. Similarly, sub-Tg transitions of tween laboratory based annealing processes (in vitro
starches (which probably represent enthalpy relaxation annealing) and environmentally driven reorganisations
[51]) have been described by other authors [51,52]. (in vivo ‘annealing’) of starch granule architecture
[23,53,57,58].
Hence, apart from species and cultivar specific varia-
tion in starch physico-chemical properties, there is a
4. Environmental considerations
profound environmental effect on gelatinisation charac-
teristics. This effect is, in the experience of these au-
The effect of environmental temperature on starch
thors, far greater than cultivar specific (and hence
synthesis and properties has been the subject of much
genetic) variation. The implications are, that product
recent research. Apart from direct growth temperature
quality is not simply a cultivar specific trait — but is
effects on the activity of enzymes involved in starch
largely dependent on environmental conditions experi-
biosynthesis (which are discussed in some detail in a
enced during starch deposition.
recent publication by these authors [53]), there is a
distinct effect on starch physico-chemical properties. In
general, for mature cereal and tuber starches [54–58]
there tends to be a small effect on the fine structure 5. Molecular basis for annealing
(chain length distribution) of amylose or amylopectin.
Granule size decreases as growth temperature increases, It has been difficult to define, at the molecular level,
while amylose content remains approximately the same. what happens to the internal structure of starch gran-
In the case of lipid (lysophospholipids and free fatty ules when they are annealed. Some authors have dis-
acids) in cereal starches (only), there is a distinct tem- cussed the molecular event in terms of increasing
perature dependent increase. Growth temperature also granule stability [32], reorganising granule structure
causes a distinct increase in gelatinisation temperatures [39,40] or lowering free energy [17]. These descriptive
8 R.F. Tester, S.J.J. Debon / International Journal of Biological Macromolecules 27 (2000) 1–12

terms do not, however, give readers a clear molecular during annealing and gelatinisation may be supported
picture associated with the reorganisations involved by the use of scanning electron microscopy [65], where
within granules when they are annealed. A number of dimensions of amorphous and crystalline lamellae may
authors have discussed annealing with more emphasis be estimated.
on the crystalline and amorphous domains. Crys- Overall, the NMR and WAXS data support the same
tallinity and crystalline ‘perfection’ (optimisation of general picture that annealing causes no significant
crystalline order) have been discussed in detail in this increase in crystalline material formed within starch
context [28,36,38,42,43,59]. Similarly, granular reorgan- granules by either of two possible mechanisms: (i)
isations have been discussed in terms of rigidity [33] formation of double helices (which need not necessarily
and realignments and partial melting [45,60]. Others be associated with existing crystalline domains) or; (ii)
recognise the importance of interactions between, and major increase in amount of crystallinity as a conse-
mobility of, amorphous and crystalline regions [3,61] quence of ordering of previously amorphous regions.
and the constituent amylose and amylopectin molecules Rather, the enhanced ordering of double helices, due to
[37]. improved registration (alignment), with associated in-
Tester et al. [23] (working on wheat starches), have creased rigidity of amorphous regions, probably under-
discussed annealing in the context of hydration and lies the annealing process.
swelling of amorphous regions (temperature range be- Unlike WAXS which quantifies crystalline order
tween Tg and To), which facilitates ordering of double throughout starch granules, small angle X-ray scatter-
helices in crystalline regions. This ordering of double ing (SAXS) quantifies differences (periodicity) at the
helices could be associated with minor optimisation of level of amorphous-crystalline lamellae radiating from
double helix length, although no additional double the hilum to the periphery of starch granules. Using
helical material is formed [23]. The amorphous material this technique, Jacobs et al. [36] showed that (for wheat
post-annealing probably becomes more ‘glassy’ (more and potato starches) the repeat distances of the crys-
rigid and less mobile) whereupon Tg is elevated. The talline and amorphous lamellae remain unchanged
constancy of double helix content pre- and post-anneal- (10.5 nm in wheat and 9.9 nm in potato), although
ing has also been shown by Jacobs [1] for a range of there was an increase in peak intensity. A pictorial
starches (pea, potato and wheat). representation of the length scales within starch gran-
With respect to the effects of annealing on the double ules together with techniques used for their quantifica-
helix content of starches, the situation in amylomaize is tion are presented in Fig. 5. More detailed discussion
far more complex than for waxy or normal starches. In regarding the application of this technique to under-
amylomaize starch there is evidence from NMR that stand structural, gelatinisation and annealing mecha-
amylose also forms some double helices and that upon nisms of starches can be found elsewhere [65]. Those
annealing there is partitioning of amylopectin and amy- authors, however, reported that the lamellar repeat
lose helical structures [62,63]. This different structural distance for wheat starch is smaller (8.85 nm) than the
model within amylomaize starch granules helps to ex- figures quoted by Jacobs et al.
plain the characteristic gelatinisation characteristics of Native cereal starch granules contain amylose–lipid
these starches (below). complexes, as shown by NMR [67,68]. The lipid
Whilst NMR can be used to quantify the number of (lysophospholipid or free fatty acid) is immobilised
double helices within starch granules, it does not mea- within the a-glucan helices and the corresponding lack
sure crystallinity per se. For this purpose, wide angle of mobility of methylene groups can be determined
X-ray scattering (WAXS) may be employed. Early using this technique. The biochemical significance of
work on annealing of wheat starch indicated that there these complexes is uncertain, although they have a
was little detectable effect on the X-ray diffraction significant effect on starch functionality. Whilst in
pattern [31]. This has been confirmed for potato starch model amylose–lipid complex systems annealing can be
[3]. However, other workers have reported a small induced [69] because of the relative ease of mobility,
increase in intensity of the diffraction pattern (but with annealing between Tg of the starch and To of amy-
little or no effect on d-line spacings) for wheat, oat, lopectin in wheat starch has little effect on this amy-
lentil [32,64] and barley starches [46] but with a de- lose–lipid endotherm [34,35]. The effect of annealing at
crease in intensity for potato starch [32]. It is very these temperatures (e.g. 35–50°C) on amylose–lipid
useful to link together both NMR and WAXS data for complexes is predictably very unlikely, because the
starches, as they measure different levels of order. One temperature is far too low. The peak transition temper-
could, for example, have non-crystalline double helices ature of these complexes is of the order of 95–115°C.
(outside crystalline domains) within starch granules At temperatures where complexes have been annealed
that give a strong NMR signal but not a WAXS (for example 80°C, [69]) the starch would be fully
diffraction pattern. The relative significance of these gelatinised in unlimiting water. Probably Tg for the
techniques for determining starch structure and order complexes under these conditions is quite close to this
R.F. Tester, S.J.J. Debon / International Journal of Biological Macromolecules 27 (2000) 1–12 9

Fig. 5. Pictorial representation of the length scales within the starch granule together with techniques used to characterise the structural features
(adapted from [66]).

temperature, although it is perhaps as mobile as Tg for steric hindrance is diminished as a consequence of the
amylopectin as a function of moisture content [23]. increased mobility during annealing. Chemically intro-
There is a relationship between the amount of starch duced phosphate groups have similar effects to natu-
phosphorylation and elevation of gelatinisation temper- rally (during biosynthesis) inserted groups [71]. This
atures as a consequence of annealing [70]. The shift in should be viewed in the context of phosphate esters
Tp (or DTp) is greater for starches with low levels of affecting the crystallinity of native starches [72], where
phosphorylation. The authors [70] proposed that this is the gelatinisation enthalpy is inversely related to the
because the number of potential dislocations is smaller. phosphate content.
In other words, phosphate moieties restrict double helix
(and consequently crystallite) formation. The higher the
phosphate content, the greater the interference. Because 6. Physical consequences of annealing
of the detrimental effect of phosphate groups on crys-
tallite formation, however, the increase in enthalpy is According to some authors [32], annealing causes no
largest for the high phosphate starches indicating that effect on granule dimensions or shapes, although early
10 R.F. Tester, S.J.J. Debon / International Journal of Biological Macromolecules 27 (2000) 1–12

microscopic work indicated that wheat starch granule (amorphous) lipid free amylose (FAM) according to the
dimensions increase after annealing [31]. Clearly, how- definition of Morrison et al. [68], the amylose must be
ever, small differences in size cannot be accurately more restricted from leaching out of the granules. Al-
quantified using microscopy and care should be placed though at a given temperature post-annealing, the gran-
on reliance upon this data. The A-type diffraction ules will swell less than un-annealed starches and this
pattern of wheat starch is retained after annealing [31], will be the primary restraint to leaching. This does,
although the line intensity may increase as discussed however, strengthen the view that there is molecular
above. Although heat moisture treatments causes a B- reorientation in the starch granule which makes the
to A-type transition for potato starches, this does not amorphous material more glassy with an elevated Tg.
happen during annealing [3]. Hence, the molecular re-
orientation is more subtle.
The effect of annealing on gelatinisation characteris- 8. Chemical hydrolysis
tics is well established, particularly using DSC, where
there tends to be an increase in To and Tp, decrease in Annealing tends to reduce the amount of acid hy-
the gelatinisation range (Tc – To) and either constancy or drolysis of starch granules, although small granules
an increase in gelatinisation enthalpy sometimes exhibit little difference or even enhanced
[3,23,28,31,32,34,37 – 41,43,46,64,70,71,73]. The increase hydrolysis [32,34]. This discrepancy has to some extent
in gelatinisation temperatures is associated with a de- been resolved by Tester et al. [23] who investigated acid
crease in swelling power [23,32,41,46,73], provided that hydrolysis for native and annealed wheat starch as a
some granular structure is retained. This is reflected in function of time. They found that for the first phase of
a higher temperature onset of swelling and reduced acid hydrolysis (0–7 days, representing amorphous ma-
swollen volume (below circa 90°C, provided that water terial hydrolysis) annealed starch was more extensively
is not limiting). The effects of annealing on pasting hydrolysed than native starch. After 7 days, where
characteristics are complex. In some studies the consis- crystalline material is progressively hydrolysed, the ex-
tency (viscosity) of annealed starches (wheat and tent of hydrolysis for the native and annealed starches
potato) increases (with associated decrease of peak was essentially the same. The explanation for the en-
viscosity for potato starch) while for lentil and oat it hanced hydrolysis of amorphous regions after anneal-
tends to decrease [32]. Similar results have been re- ing is that the amorphous regions become more
ported for wheat and potato starch, with annealed pea concentrated due to the enhanced glassy structure. On
and rice starches exhibiting increased viscosity [3,33,41]. the other hand, the similarity in hydrolysis pattern
Using the model proposed by these authors and during the crystalline hydrolysis phase (\ 7 days) confi-
co-workers [23], the physical properties discussed above rms that it is double helices (which remain constant)
can be explained on the basis of more glassy amor- which are the primary contributor to the hydrolysis
phous regions within annealed starch granules and a profile during this phase. Amylose–lipid complexes
more ordered registration of amylopectin double he- may affect the pattern of acid hydrolysis in cereal
lices. These molecular events restrict ease of hydration starches, which could also influence enzymic hydrolysis
of the starch granules during gelatinisation and elevate (below).
gelatinisation temperatures. In parallel, these events
restrict swelling. It is difficult to unravel the effects on
pasting characteristics, because this system will be 9. Enzymatic hydrolysis
strongly influenced by granule size and polysaccharide
solubilisation — more so than gelatinisation (by DSC) Certain studies have indicated that annealed wheat,
and swelling power determinations. This is probably barley and sago starches are more easily hydrolysed by
why there is a lack of consistency in response to a-amylases than native starches [31,46,74]. This has,
annealing for starches from different botanical origins. however, been contradicted by other research on wheat,
lentil and potato starches [32,41], although small starch
granules (oat) have been reported to be much more
7. Solubility easily hydrolysed post-annealing [32]. However, the rate
of a-amylase hydrolysis for different starches follows
The annealing process itself leads to little solubilisa- two distinct phases: an initial rapid then subsequent
tion of a-glucan [23]. This is important as it shows that slow phase. Annealing alters the extent of hydrolysis of
improved order is a genuine molecular event rather these different phases as a function of botanical origin
than a consequence of leaching amorphous a-glucan [35]. During the second phase of hydrolysis, annealed
and hence ‘concentrating’ crystalline material. Anneal- wheat and pea starches are more resistant to a-amylase
ing reduces solubilisation of a-glucan during swelling hydrolysis whilst the inverse is true for potato starch
below 100°C [32,41,46,73]. As leachate is primarily [35]. Annealed potato starch is less easily hydrolysed by
R.F. Tester, S.J.J. Debon / International Journal of Biological Macromolecules 27 (2000) 1–12 11

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