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Nuclear Instruments and Methods in Physics Research B 265 (2007) 399405

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DSC studies of retrogradation and amyloselipid complex

transition taking place in gamma irradiated wheat starch
K. Ciesla
b

a,*

, A.C. Eliasson

a
Institute of Nuclear Chemistry and Technology, Dorodna 16 str., 03-195 Warsaw, Poland
Department of Food Technology Engineering and Nutrition, Division of Food Technology, University of Lund, P.O. Box 124, S-221 00 Lund, Sweden

Available online 8 September 2007

Abstract
The eect of gamma irradiation (60Co) with doses of 530 kGy on the amyloselipid complex transition and retrogradation occurring
in gels containing ca. 50% and ca. 20% wheat starch was studied by dierential scanning calorimetry (DSC) during heatingcoolingheating cycles (up to three cycles). Transition of the amyloselipid complex occurs in all the irradiated samples at a lower temperature as
compared to the non-irradiated starch. That eect was larger when the radiation dose was higher. A further thermal treatment causes
a decrease of the transition temperature in the irradiated samples, with no eect or increase of that temperature observed for the nonirradiated ones. Irradiation hinders retrogradation taking place in 50% gels but facilitates the process occurring in 20% gels. The dierences between the irradiated and the non-irradiated samples are more evident in the every next heating or cooling cycle as well as after
storage and in the case of ca. 50% suspensions as compared to ca. 20% suspensions. The results point out to the deterioration of the
structure of the complexes formed in the irradiated starch as compared to the non-irradiated one.
2007 Elsevier B.V. All rights reserved.
PACS: 81.40.Gh; 81.40.Wx; 81.70.Pg; 87.14.Df; 87.15.Zg; 61.82.Pv
Keywords: Wheat starch; Gamma irradiation; Gelatinisation; Amyloselipid complex transition; Retrogradation; Dierential scanning calorimetry (DSC)

1. Introduction
During the last decade interest has increased in the
methods of food sterilisation and modication applying
medium (110 kGy) and high doses (1070 kGy) of irradiation [16] as well as radiation processing of industrial
products that contain starch. Foods sterilised at high doses
may be consumed by immunologically depressed patients
and can be stored at room temperature (for example bakery products, readily prepared meals). Radiation modication enables, moreover, removal of antinutritional factors
and inhibition of food allergies [36]. Doses of several
dozen kGy are used for sterilisation of pharmaceuticals
and medical devices and for starch modication. Accordingly, it appears desirable to acquire knowledge about the

Corresponding author. Tel.: +48 22 5041106; fax: +48 22 8111917.

E-mail address: kciesla@orange.ichtj.waw.pl (K. Ciesla).

0168-583X/$ - see front matter 2007 Elsevier B.V. All rights reserved.
doi:10.1016/j.nimb.2007.09.010

functional and structural properties of foods and starch

alone irradiated using medium and high doses and in the
development of appropriate physico-chemical testing methods. A good recognition of radiation inuence on the properties of wheat starch is of great importance, since it
constitutes a component of a number of food products.
Simultaneously, there is a lack of data concerning the eect
of ionising radiation on starchlipid and starchsurfactant
interactions (apart to our previous work [710]), while such
systems are examined intensively in relation to food, pharmaceutical and other industries. The relationship between
the structural properties of starchlipid (surfactant) complexes and the digestibility of food is still the subject of
studies.
Dierential scanning calorimetry (DSC) is a valuable
method for obtaining information about starch properties.
Gelatinisation and Amyloselipid complex transition
occurring on heating of starch and our suspensions as well
as retrogradation taking place during the further storage of

400

K. Ciesla, A.C. Eliasson / Nucl. Instr. and Meth. in Phys. Res. B 265 (2007) 399405

the resulting gels depend on the molecular structure of

polysaccharides chains and their ordering in starch granules. The interaction of starch with lipids inuences gelatinisation and hinders unprotable recrystallisation of gels
[11,12]. Formation of starchlipid complexes occurs during
gelatinisation taking place on heating of starch suspensions
at temperatures above ca. 45 C. The inclusion helical complex formed between lipids and amylose chains undergoes a
partially reversible transition (of the order/disorder type) at
higher temperature. Endothermal eects of the complexes
melting are observed during heating at temperature above
80 C, while exothermal eects of crystallisation occur during cooling. There is no evidence, however, for thermal
eects connected to the formation of lipid complexes with
the branched amylopectin macromolecules.
Our previous studies (conducted using WAXS, SAXS
and DSC methods) have shown that degradation brought
about by gamma irradiation [13] induces a decrease in
the order of starch granules [14,15] and consequently inuences gelatinisation [710,16]. It was also found that modication of lipids surrounding and lipids themselves
resulting from the irradiation aect the properties of
starchlipid complexes formed in wheat starch and wheat
our [69]. In particular, the smaller enthalpies of gelatinisation of Amyloselipid complex transition were determined in the cases of the starches irradiated with a dose
of 30 kGy as compared to the non-irradiated ones. An
essential decrease in the transition temperature was also
found after irradiation. These results were attributed to a
deterioration of the complex structure resulting due to irradiation. Moreover, the additional deterioration of the complex symmetry could be concluded after the further thermal
treatment of the irradiated gels, on the basis of deterioration of thermal eects and their shift to lower temperature
at each subsequent heating or cooling cycle. Simultaneously, the increase in the symmetry of the complex
formed in the non-irradiated starch was concluded after
the same thermal treatment. Furthermore, our preliminary
results have demonstrated that dierences in the storage
eects on the irradiated and non-irradiated wheat starch
and our gels might result in the expanded dierences
between the structures of Starchlipid complexes formed
in such gels. It was found that the degree of retrogradation
in the 50% gels of wheat starch was smaller after irradiation with a 30 kGy dose, while the degree of retrogradation
of ca. 20% wheat our was higher as compared to the
respective non-irradiated specimens stored at the same
conditions.
The studies carried out until now concerned the inuence of irradiation with the dose of 30 kGy. Moreover,
the studies of the storage eect on retrogradation and
Amyloselipid complex structure had only a preliminary
character. The purpose of the present detailed study was
to determine the eect of irradiation carried out using
doses in the range from 5 to 30 kGy on the properties of
Amyloselipid complex formed in wheat starch as well as
on retrogradation taking place in the gels. The inuence

of thermal treatment connected to the DSC measurements

and eect of the further storage on the possibility to
observe dierences between transition of the Amyloselipid
complex occurring in the non-irradiated and the irradiated
starch gels was tested. The radiation eects on the processes occurring in the gels characterised by the primary
dry matter to water ratio of 1:1 and 1:4 were studied.
Terms dense and watery suspensions/gels are used in the
next paragraphs in relation to these systems, respectively.
2. Experimental
Solid native wheat starch of Sigma production (S-5127)
containing ca. 11.7 wt% of water was irradiated with 60Co
gamma rays with doses of 5, 10, 20 and 30 kGy applying a
dose rate of 1.00 0.05 Gy s 1. Irradiations were carried
out in air at room temperature in a gamma cell Issledovatiel, installed in the Department of Radiation Chemistry in
the Institute of Nuclear Chemistry and Technology, Warsaw, Poland.
DSC studies were carried out in an inert gas stream
(nitrogen) during heatingcoolingheating cycles (up to
three heating/cooling processes) within the temperature
range of 5150 C. A Seiko DSC 6200 calorimeter was
used, operating at heating and cooling rates of
10 C min 1. The instrument was calibrated with gallium
(Mp = 29.8 C) and indium (Mp = 156.6 C). Covered aluminium pans from TA Instruments (20 lL large) were used
in the experiments and a pan with aluminium oxide was
used as the reference. Portions of starch (ca. 2.2 mg and
ca. 4.2 mg in purpose to prepare ca. 20% and ca. 50%
suspensions, respectively) were weighted directly in the
pre-weighted DSC pans and the required amounts of twice
distilled water were added. The pans were then hermetically
closed and re-weighed. The residues obtained after the rst
DSC analyses were stored at 4 C for 7 days (in the case of
watery samples) or at ambient temperature for 13 days
(dense suspensions) and then the analyses were repeated
applying the single heating/cooling cycle. The sample pans
Table 1
The values of peak temperature (Tp, C) determined for thermal eects of
the transition of Amyloselipid complex taking place during heating
Dose (kGy)

Heating cycle (C)

II

III

Dense gels (48.3150.01%)

0
112.1 0.3
5
112.5 0.1
10
112.6 0.2
20
112.7 0.2
30
112.2 0.2

116.1 0.2
115.7 0.1
114.9 0.3
113.9 0.2
113.3 0.2

116.2 0.2
115.6 0.2
114.6 0.2
113.4 0.1
112.1 0.3

Watery gels (21.9823.78%)

0
99.0 0.1
5
98.9 0.5
10
98.9 0.4
20
97.5 0.4
30
97.3 0.5

101.9 0.5
101.5 0.7
101.3 0.5
100.7 0.5
100.3 0.1

102.0 0.5
101.4 0.7
100.7 0.5
100.2 0.5
99.5 0.3

K. Ciesla, A.C. Eliasson / Nucl. Instr. and Meth. in Phys. Res. B 265 (2007) 399405

401

Table 2
The values of peak and onset temperature determined for thermal eects of the transition of Amyloselipid complex taking place during cooling
Dose (kGy)

Cooling cycle (C)

I
Tp

Dense gels (48.3150.01%)

0
99.7 0.2
5
99.5 0.1
10
98.8 0.1
20
97.8 0.1
30
95.8 0.5
Watery gels (21.9823.78%)
0
83.5 0.1
5
83.3 0.2
10
83.0 0.1
20
82.3 0.1
30
81.6 0.1

II
Ton
101.5
101.5
100.7
99.8
98.2

III

Tp
0.2
0.1
0.1
0.1
0.3

85.6 0.1
85.1 0.5
84.9 0.2
84.1 0.3
83.1 0.1

Ton

Tp

Ton

0.3
0.2
0.1
0.1
0.2

101.7 0.3
101.4 0.2
100.5 0.1
98.4 0.0
97.3 0.2

100.2 0.4
99.4 0.3
98.0 0.4
95.9 0.1
94.8 0.3

102.0 0.4
101.7 0.2
100.2 0.2
97.8 0.3
96.8 0.2

83.5 0.1
83.0 0.2
82.5 0 .1
81.7 0.1
80.8 0.2

83.6 0.1
84.9 0.2
84.2 0.2
84.1 0.2
82.6 0.2

83.5 0.1
82.7 0.2
82.0 0.1
81.4 0.1
80.2 0.1

85.7 0.2
84.8 0.3
83.4 0.2
83.1 0.2
82.4 0.1

99.9
99.6
98.5
96.4
95.3

mass was controlled directly before and after the rst as

well as directly before and after the second experimental
step. We analysed the data only for which no valid decrease
in the sample mass was detected after heating in the DSC
instrument. Nevertheless, some decrease in water content
was noticed after storage. The dry matter content of each
individual sample was determined by drying punctured
DSC pans in a heating cabinet at 105 C for 16 h after all
analyses. The average values of transition enthalpy are
given in terms of dry matter content.
Peak (Tp) and onset (Ton) temperatures of thermal eects
corresponding to Amyloselipid complex as well as enthalpies of Amyloselipid complex transition (DH), and of gelatinisation (DHg) were calculated basing on several duplicate
measurements (36). In purpose to demonstrate the radiation eect on the complex Tp and Ton values were specied
in Tables 1 and 2 for thermal eect recorded at cooling and
Tp values were reported for those recorded at heating. This
is because that the similar DH values were determined for
the non-irradiated starch and the samples irradiated with
doses till 20 kGy on the basis of exothermal eects recorded
at cooling and because of the diused beginning of endothermal eects recorded at heating (Fig. 2).
In purpose of direct comparison of irradiation eect on
retrogradation, the yield of retrogradation R was calculated. This parameter expressed enthalpy of gelatinisation
found after retrogradation as a percentage of the initial
enthalpy of gelatinisation occurring during the rst heating
of the respective sample (thus taking into account enthalpy
decrease brought about by irradiation).
Several modes were tried for calculation of enthalpy
basing each of experimental set with slightly modied integration limits and the nal results were than determined as
the average values. The average variances of the results
were determined on the basis of the variances obtained
for several measurements and calculations performed using
the modied limits.
Accordingly to the fact that the processes occurring in
the presence of small water content depend strongly on

concentration, only the parameters obtained for the samples characterised by very close high concentrations were
compared.
3. Results and discussion
3.1. The primary analyses
Gelatinisation enthalpies determined for the watery
starch suspensions were all in the range of 11.4
11.0 0.3 J g 1 and in the range of 12.310.4 J g 1 in the
case of the dense suspensions (the values expected for wheat
starch). The upper limit of average values concerns the nonirradiated starch and the lower limit concerns that irradiated with a dose of 30 kGy. Integrations were done within
the similar temperature ranges for all samples: 51.1
80.0 C and 46.696.6 C, respectively in the case of watery
and dense suspensions (Fig. 1). Detailed DSC studies concerning gamma radiation inuence on gelatinisation of
wheat starch occurring in both systems are presented in [10].
No regularity could be stated between the temperature
of the complex transition taking place during the rst heating of the dense suspensions containing the specic
starch (shown by Tp values determined for the appropriate
endothermal eects; Table 1, Fig. 1). This is connected
probably to the fact, that the complex structure was not
formed yet, especially that small water content additionally
might prohibit its formation. In the case of the watery
suspensions, the lower Tp values were determined for thermal eects of the transition already during the rst heating
of the samples irradiated applying the doses of 20 and
30 kGy as compared to the non-irradiated sample (Table
1, Fig. 1). No dierence was found, however, between the
non-irradiated starch and those irradiated using the doses
of 5 and 10 kGy. The transition occurs at a higher temperature during the second heating than during the rst heating as well in dense as in watery gels, due to the
reorganisation of the complex structure resulting from
melting/recrystallisation processes.

402

K. Ciesla, A.C. Eliasson / Nucl. Instr. and Meth. in Phys. Res. B 265 (2007) 399405

Fig. 1. The examples of DSC curves recorded in the case of watery

(22.92%) and dense (49.85%) starch suspensions during the rst heating.
Thermal eects observed at respective curves at temperature range till
80.0 C and till 96.6 C correspond to gelatinisation and those observed at
higher temperature are attributed to Amyloselipid complex transition.
Discussion concerning double endothermal minima connected to gelatinisation taking place in dense system is given in [10].

All the irradiated samples revealed a decreased temperature of the transition occurring on the rst cooling and the
subsequent heating and cooling cycles (shown by the lower
average Tp and Ton values) as compared to the non-irradiated starch. This could be concluded already after irradiation with a dose of 5 kGy. Moreover, the respective Tp and
Ton values were the lower when the irradiation doses were
the higher (Tables 1 and 2). A smaller height and diused
borders of thermal eects detected for the irradiated starch
both on heating and on cooling as compared to those
recorded for the non-irradiated starch (Fig. 2), together
with the decreased temperature of the transition, point
out to a decrease in the ordering of the complex.
Thermal treatment connected to DSC measurements
causes a decrease of the transition temperature in the irradiated dense as well as watery gels (showing the further
deterioration of the complex structure), with an exception
of no eect on the dense gels of starch irradiated with a
dose of 5 kGy (Fig. 3, Tables 1 and 2). Simultaneously,
no eect or increase of transition temperature was observed
in the non-irradiated starch submitted to the same treatment. Accordingly to those dierences in the sensitivity
of the complexes, the dierences between all the irradiated
and the non-irradiated samples became easier seen in every
next heating or cooling cycle.
3.2. The secondary analyses
Broad thermal eects observed during heating above
40 C result due to gelatinisation of the partially recrystal-

Fig. 2. Comparison of the endothermal eects recorded during the third

heating and exothermal eects of the Amyloselipid transition observed
during the third cooling in the case of the dense systems: 0 kGy 50.01%
and 30 kGy 48.31%.

Fig. 3. The examples of DSC curves recorded during the rst, the second
and the third cooling dense gels, non-irradiated (48.31%) and irradiated
using a dose of 30 kGy (49.73%).

lised gels (Fig. 4; compare to Fig. 1, curve 2 recorded at the

rst heating). Irradiation hinders retrogradation taking
place in dense suspensions (Fig. 4, Table 3) similarly as
in the case of wheat our [10]. This is shown by the
decreased enthalpy of gelatinisation and the yield of retrogradation R determined after storage of the irradiated samples as compared to the non-irradiated starch.
Simultaneously, retrogradation taking place in the watery
gels was facilitated after irradiation (Table 3), as it was stated previously after irradiation performed with a 30 kGy
dose [8].
During the heating cycle carried out after retrogradation
of the dense suspensions, the transition was observed at a

K. Ciesla, A.C. Eliasson / Nucl. Instr. and Meth. in Phys. Res. B 265 (2007) 399405

Fig. 4. The examples of DSC curves recorded after retrogradation of

dense gels (room temperature, 13 days): 0 kGy (60.51%), 20 kGy (61.09%)
and 30 kGy (60.91%).

lower temperature when irradiation dose was higher (starting from 127.6 0.5 C in the case of the non-irradiated
sample; Table 3). However, when the starch was irradiated
with a dose of 20 kGy, additional endothermal peaks preceded the major endothermal eect recorded at only
slightly lower temperature than in the case of the starch
irradiated with a dose of 10 kGy. Peaks of these thermal
eects were observed in the wide temperature range of

403

94110 C (Fig 2, Table 3). When the radiation dose reach

30 kGy, these new peaks became more evident and larger
than the thermal eects observed in the temperature range
above 120 C. The appearance of these additional transitions can be attributed to the separation of the complex
form with a lower symmetry resulting due to the random
distribution of starchlipid helices [17]. However, multiple
minima on DSC curves might result also due to the occurrence of the recrystallisation during melting of the complex
characterised by the deteriorated structure. These explanations both indicate, however, the existence of the complex
form with a decreased symmetry in that irradiated and retrograded starch as compared to the retrograded non-irradiated starch.
Deterioration of the complex structure connected both
to irradiation and to storage can be also concluded on
the basis of the essentially diused single thermal eects
recorded during cooling of dense gels (Fig. 4). These eects
were more blurred after retrogradation than at the primary
analysis and the dierences between the heights of the
peaks recorded for the non-irradiated and the irradiated
samples were expanded (compare Fig. 2 and Fig. 4) as well
as the dierences between Tp and Ton values (Tables 24).
Decrease in enthalpy of the complex transition occurring
on cooling became evident and detected already after irradiation with a dose of 5 kGy (Table 3) while no change in
enthalpy was found during the primary analysis due to
irradiation with that dose (Table 2). The smaller enthalpies
found for the irradiated starches might be connected to the
limited binding of lipids. A blurred beginning of thermal
eects suggests, however, that a large fraction of complexes
characterised by the low ordering underwent transition in a
wide range of lower temperature, without the well dened
thermal eects.
In the case of transition taking place in the recrystallised
watery gels during heating as well as cooling, the similar

Table 3
Results of the repeated DSC analyses performed for the residues obtained after the rst analyses described in Tables 1 and 2
Dose(kGy)

Retrogradation
DHg

(J g 1)

Amyloselipid complex transition

R (%)

Heating

Cooling

Tp (C)

Tp (C)

Ton (C)

111.0 0.5
109.1 0.1
106.8 0.5
105.7 0.3
104.2 0.8

112.7 0.8
111.3 0.2
109.7 0.9
108.2 0.1
107.3 1.0

1.93 0.10
1.53 0.13
1.35 0.06
1.09 0.15
0.71 0.15

84.3 0.2
83.5 0.5
82.3 0.3
82.2 0.3
79.9 0.1

86.5 0.2
85.1 0.3
84.1 0.3
84.0 0.3
82.3 0.1

1.84 0.10
1.64 0.30
1.90 0.10
1.79 0.10
1.50 0.06

Dense gels (58.6061.09%), 13 days of storage at room temperature

0
9.53 0.45
80
127.6 0.5
5
8.30 0.13
72
125.3 0.0
10
7.60 0.90
67
124.2 0.8
20
6.37 0.17
60
123.8 0.8b and 94.7111.8c
30
5.91 0.30
54
97.2110.8b and 120.0123.2c
Watery gels (24.6527.16%) 7 days of storage at 4 C
0
60.50
64
5
0.70 0.30
6
10
1.10 0.30
9
20
2.00 0.30
17
30
2.70 0.30
25
a
b
c

103.0 0.3
102.5 0.5
101.1 0.3
100.3 0.3
99.5 0.3

Enthalpy was calculated in the range of 39.386.6 C.

Two or three endothermal eects were observed in particular thermograms, Tp value of the major thermal eect is underlined.
Due to weak reproducity of the additional peaks positions, the range of possible occurrence was shown.

DH (J g 1)

404

Dose (kGy)

R
determined after storage (%)

Tp (0 kGy)Tp (C)
Heating
Primary analysis,
the third cycle

Cooling
Repeated analysis

Primary analysis,
the third cycle

Repeated analysis

Dense gels (48.3150.01% at the primary analysis and 58.6061.09% at the analysis repeated after 13 days of storage)
0
80
0.0
0.0
5
72
0.6
2.3
10
67
1.6
3.4
20
60
2.8
3.8a
30
54
4.1
nc

0.0
0.8
2.2
4.3
5.4

0.0
1.9
4.2
4.6
6.8

Watery gels (21.9823.78% at the primary analysis and 24.6527.16% at the analysis repeated after 7 days of storage
0
64
0
0
5
6
0.6
0.3
10
9
1.3
1.9
20
17
1.8
2.7
30
25
2.5
3.5

0
0.8
1.5
2.1
3.3

0
1.4
2.4
2.5
4.2

Tp (0 kGy)Tp is the dierence between Tp value determined for the non-irradiated sample and the appropriate value determined for the sample irradiated using the respective dose.
nc not calculated.
a
The major eect of the multiple melting occurring in the retrograded samples were taken into account.

K. Ciesla, A.C. Eliasson / Nucl. Instr. and Meth. in Phys. Res. B 265 (2007) 399405

Table 4
Comparison of the possibilities to observe the dierences between Tp values determined in the case of the non-irradiated and the specic irradiated samples during the third heating and cooling cycles of
the primary analysis with those obtained after storage

K. Ciesla, A.C. Eliasson / Nucl. Instr. and Meth. in Phys. Res. B 265 (2007) 399405

statements can be done as those derived from the analyses

performed for dense gels during cooling.
The increase in the temperature of the complex transition after storage in relation to the last cycle of the rst
analysis was connected partially to the increase in concentration. However, the increase in Tp was larger in the case
of the non-irradiated starch than in the case of the irradiated ones observed at strictly the same conditions. This
occurred in both dense and watery systems, therefore
independently whether retrogradation was restricted or
facilitated due to irradiation. For example, the dierence
between Tp values recorded in those two cycles were
equal to 10.8 and 9.4 in the cases of dense gels, respectively non-irradiated and that irradiated with a 30 kGy
dose.

405

Irradiation facilitates retrogradation taking place in

the watery system but hinders the process occurring in
the dense system. Therefore, irradiation might appear the
appropriate treatment for obtaining more stable dense gels
and pastes, and consequently for improving the properties
of ready-to-eat meals as well as some industrial products.
Results concerning the radiation eect on the structure
of starchlipid complexes in wheat starch might be related
to the starch/lipid and starch/surfactant systems that found
application in food, pharmaceutical and other industries.
Acknowledgement
The work was done in the frame of Research Grant 2
P06T 026 27 of Polish Ministry of Science and Higher
Education.

4. Conclusion
References
Modication of the Amyloselipid structure in wheat
starch, in particular decrease of the complex symmetry,
can be concluded after irradiation with doses of 5
30 kGy. Melting and crystallisation occurring under thermal treatment and storage in the non-irradiated starch
and the starches irradiated using various doses induces various modication of starchlipid complexes. Deterioration
in ordering of the strongly irradiated complexes can be stated after the further treatment of gels while improvement of
the structure might be concluded in the case of non-irradiated starch and the intermediate eects are noticed after
irradiation performed with the intermediate doses. Dierences between the irradiated and the non-irradiated samples are more evident in the every next heating or cooling
cycle as well as after storage and in the case of dense gels
(characterised by starch:water ratio equal to 1:1) as compared to the watery ones (starch:water ratio equal to 1:4).
These observations might appear useful for possible detection of irradiation. In particular, Tp values of thermal eect
of the Amyloselipid complex transition, decreasing in the
every subsequent heating/cooling cycle, might serve as indicator of the formerly performed irradiation.

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