ISSN 0101-2061
Original
Ciência e Tecnologia de Alimentos
he efect of acid hydrolysis on the technological functional
properties of pinhão (Araucaria brasiliensis) starch
Efeito da hidrólise ácida nas propriedades funcionais tecnológicas do amido de pinhão (Araucaria brasiliensis)
Roberta Cruz Silveira THYS1*, Andréia Gomes AIRES1,
Ligia Damasceno Ferreira MARCZAK2, Caciano Pelayo Zapata NOREÑA1
Abstract
Technological functional properties of native and acid-thinned pinhão (seeds of Araucária angustifolia, Brazilian pine) starches were evaluated
and compared to those of native and acid-thinned corn starches. he starches were hydrolyzed (3.2 mol.L–1 HCl, 44 °C, 6 hours) and evaluated
before and ater the hydrolysis reaction in terms of formation, melting point and thermo-reversibility of gel starches, retrogradation (in a
30-day period and measurements every three days), paste freezing and thawing stability (ater six freezing and thawing cycles), swelling
power, and solubility. he results of light transmittance (%) of pastes of native and acid-thinned pinhão starches was higher (lower tendency
to retrogradation) than that obtained for corn starches ater similar storage period. Native pinhão starch (NPS) presented lower syneresis
than native corn starch (NCS) when submitted to freeze-thaw cycles. he acid hydrolysis increased the syneresis of the two native varieties
under storage at 5 °C and ater freezing and thawing cycles.
he solubility of NPS was lower than that of native corn starch at 25, 50, and 70 °C. However, for the acid-thinned pinhão starch (APS), this
property was signiicantly higher (p < 0.05) when compared to that of acid-thinned corn starch (ACS). From the results obtained, it can be
said that the acid treatment was eicient in producing a potential fat substitute from pinhão starch variety, but this ability must be further
investigated.
Keywords: unconventional starch source; modiied starch; fat substitutes.
Resumo
As propriedades funcionais tecnológicas do amido nativo e modiicado (hidrólise ácida) de pinhão (Araucaria angustifólia) foram comparadas
às propriedades do amido nativo e ácido hidrolisado de milho. As espécies de amido foram hidrolisadas (3.2 mol.L–1 HCl, 44 °C, 6 horas)
e avaliadas, antes e após a reação de hidrólise, de acordo com as análises de formação, fusão e termorreversão do gel, retrogradação (em
um período de 30 dias, com medidas a cada 3 dias), estabilidade ao congelamento e descongelamento (após 6 ciclos de congelamento e
descongelamento), poder de inchamento e índice de solubilidade. Os resultados obtidos demonstraram que o amido de pinhão apresenta
menor tendência à retrogradação quando comparado ao amido de milho, tanto para a forma nativa quanto na modiicada, após períodos
similares de armazenamento. O amido nativo de pinhão (APN), quando submetido a sucessivos ciclos de congelamento e descongelamento,
apresentou menor sinérese do que o amido de milho nativo (AMN). Nas temperaturas de 25, 50 e 70 °C, a solubilidade do APN foi menor do
que a obtida pelo AMN. Entretanto, para a forma modiicada, o amido de pinhão apresentou maior solubilidade (p < 0,05) do que o amido de
milho. Através dos resultados, pode-se airmar que o tratamento ácido modiicado realizado no amido de pinhão foi efetivo para a produção
de um potencial substituto de gordura, propriedade que deve ser testada e analisada em estudos futuros.
Palavras-chave: fonte de amido não convencional; amido modiicado; substituto de gordura.
1 Introduction
Brazilian Pine (Araucaria brasiliensis syn. A. angustifolia)
belongs to the Araucariaceae family and is the most economically
important native conifer species in Brazil (ZANDAVALLI;
DILLENBURG; DE SOUZA, 2004). The seed of this tree,
harvested from April to August, is known as pinhão, and it is
most commonly eaten ater being cooked and peeled. Pinhão is
also used as raw lour as an ingredient for several dishes, and is
considered a source of starch (~36%), dietary iber, magnesium
and copper, besides producing a low glycemic index ater its
consume (CORDENUNSI et al., 2004). Although nutritional
and technological aspects of pinhão are scarce in the scientiic
literature, recent studies suggest that the Araucaria seed is a
potential alternative source of starch extraction for industrial
purposes (CORDENUNSI et al., 2004; BELLO-PEREZ et al.,
2006; STAHL et al., 2007).
Starch is the most commonly thickening and gelling
agent used by the food industry in the development of a large
number of products such as soups, lans, sauces, and readyto-eat food among others. In recent years, there has been an
efort of researchers to ind new sources of unconventional
native starch with the necessary properties for the food
industry, such as absence of syneresis, transparency, stability,
Received 22/6/2012
Accepted 16/10/2012 (00T5756)
1
Institute of Food Science and Technology – ICTA, Federal University of Rio Grande do Sul – UFRGS, Av. Bento Gonçalves, 9500, Campus do Vale, CEP 91501-970,
Porto Alegre, RS, Brazil, e-mail: roberta.thys@ufrgs.br
2
Department of Chemical Engineering, Federal University of Rio Grande do Sul – UFRGS, CEP 91501-970, Porto Alegre, RS, Brazil
*Corresponding author
Ciênc. Tecnol. Aliment., Campinas, 33(Supl. 1): 89-94, fev. 2013
89
Evaluation of technological functional properties of acid-thinned pinhão starch
and solubility in cold water (ZHANG et al., 2005). One of the
reasons for such interest is the fact that native starch, despite
being a good texture stabilizer and regulator in food systems, has
limitations such as low resistance and thermal decomposition
and high tendency to retrogradation, which limits its use in
some industrial applications (HERMANSSON; SVEGMARK,
1996). hus, according to Flores-Gorosquera et al. (2004), it is
clear that, unlike the standard range of cereal grains (e.g. corn,
wheat, rice) and tubers (e.g. potato, cassava), alternative starch
sources are needed.
Currently, to overcome function problems of native
starches, the modiication of the polysaccharide molecule has
been a frequent practice in order to better meet market needs.
Modiied starches show better paste clarity and stability
(by oxidation), increased resistance to retrogradation, and
freeze-thaw stability (by acid hydrolysis) (BEMILLER, 1997).
Stahl et al. (2007) previously reported the modiication of pinhão
starch by phosphorylation process. However, no reports were
found on acid-thinned pinhão starch.
Starches used in gum candies and baked goods are generally
modiied with hydrochloric acid and are termed ‘‘acid-thinned’’
starches; they are obtained by chemical, physical, or enzymatic
methods (AGBOOLA; AKIMBALA; OGUNTIMEIN, 1991).
In the chemical method, hydrolyzed starches are produced
when a concentrated suspension of starch (30-40 g/100 g
solids) is treated with acid (hydrochloric acid and sulfuric acid)
at temperatures lower than those of gelatinization (30-60 °C)
for one or many hours of reaction time (FLECHE, 1985). Acid
hydrolysis reduces the molar mass, and consequently it increases
the free aldehyde group content. It also decreases viscosity,
increases the solubility of the granules, minimizes syneresis,
and causes gel thermo-reversibility when subjected to cooling
ater melting (WHISTLER; DANIEL, 1990) creating a potential
fat substitute for the food industry.
(2003). he seeds were handily uncoated; the starch recovery
from the coated seeds was 33.09%, and from the uncoated seeds
it was 42.97%.
2.3 Preparation of acid-thinned pinhão (APS) and corn
(ACS) starches
Acid-thinned starches were prepared according to the Mun
and Shin (2006) method. A total of 10g/40 mL (w/v) starch slurry
was prepared by adding aqueous HCl solution (3.2 mol.L–1) to
water bath (44 °C) with constant stirring (150 rpm). he pH
of the starch suspension ater the HCl addition was 2.8 ± 0.2.
Ater six hours, the pH of the slurry was adjusted to 5.5 ± 0.2
by slow addition of aqueous sodium hydroxide (1 mol.L–1). he
slurry was centrifuged (2000 rpm/10 min/25 °C) and washed
three times with distilled water (40 mL), followed by a new
centrifugation. he supernatant pH was adjusted to 7.5 ± 0.2
by slow addition of aqueous sodium hydroxide (1 mol.L–1)
in order to determine the dextrose equivalent (DE). The
starch was dried in a convection oven at 45 °C until reaching
moisture content of 11-13% and was ground in a hammer mill
(Fritsch – Pulverizette). For comparison purposes, corn starch
(Maizena®, Brazil) was hydrolyzed in the same conditions.
2.4 Dextrose equivalent measure (DE)
he reducing sugars content was determined by DE analysis,
carried out using dinitro-salicylic acid (DNSA) (MILLER, 1959).
he concentration of dextrose equivalent was obtained by a
standard curve previously prepared with solutions of diferent
concentrations of glucose (glucose mg/mL solution).
2.5 Formation, melting point, and thermo-reversibility of
the starch gel
2 Materials and methods
The gel formation was determined following the
methodology described by the National Starch and Chemical
Corporation (NATIONAL..., 1985). The obtained gel was
melted in a water bath at 100 °C under agitation, according
to the procedure reported by Richter, Schierbaum and KlausDieter (1973). he change in consistency was visually observed,
and the temperature at which the gel completely melted was
registered and regarded as the melting point of the gel. For
the gel thermo-reversibility, the gel was melted in a water bath
with constant stirring (Precision Scientiic Reciprocal Shaking
Bath) and allowed to cool down to room temperature, followed
by refrigeration at 5 °C for 18 hours. he formation of gel was
observed and registered (AMAYA-LLANO et al., 2008).
2.1 Materials
2.6 Light transmittance (%)
Seeds of pinhão and native corn starch (food grade) were
obtained from the local market in Porto Alegre (RS, Brazil)
between April and July (2009). All reagents were purchased
from Merck.
Light transmittance of starch paste from native and acidthinned pinhão and corn starches was measured as described
by Albrecht, Nelson and Stainberg (1960).
In order for starch to be used as fat substitute, an amylose
content of ~20% is recommended, as well as low levels of
lipids and proteins attached to the surface (VANDERVEEN;
GLINSMANN, 1992).
In a previous study, Cordenunsi et al. (2004) showed that
the native pinhão starch has interesting structural and functional
characteristics as a new source of starch; the granules had a low
protein (3%) and lipids content (1.3%) and an amylose content
of 26%. he objective of this study was to determine the efect
of chemical modiications (acid hydrolysis) on the technological
functional properties of pinhão starch.
2.7 Paste freeze-thaw stability
2.2 Pinhão starch extraction
Pinhão starch was extracted from uncoated seeds (without
the internal and external coats) by the method of Leonel et al.
90
Paste freeze-thaw stability was determined as described
by White, Abbas and Johnson (1989). Starch gel (5%, w/w dry
basis, total weight 28 g) was prepared by heating at 95 °C for
Ciênc. Tecnol. Aliment., Campinas, 33(Supl. 1): 89-94, fev. 2013
Thys et al.
30 minutes. he resulting gel was allowed to cool down to room
temperature for 15 minutes, and the gel was stored in a domestic
freezer at –5 °C over night and thawed at ambient temperature
(25 °C ± 2 °C) for 2 hours, repeatedly for up to six cycles. Ater
the last freezing and thawing cycle, the gel was centrifuged at
6000 rpm for 30 minutes, and the water separated was measured.
Stability was expressed as the percentage of water separated ater
six cycles of alternate freezing and thawing.
2.8 Swelling power and solubility
Swelling power and solubility patterns were determined
according to the method proposed by Sathe and Salunkhe (1981)
using the Equations 1 and 2, respectively:
Swelling Power (SP ) =
% solubility
weight of swollen granules ( g)
Sample weight in dry basis ( g)
dry weight at 120°C ( g)
× 100
Sample weight in dry basis ( g) × 10
(1)
(2)
2.9 Statistical analysis
he data reported in the Tables were subjected to one-way
analysis of variance (ANOVA) using Minitab Statistical Sotware
version 7.0 (Minitab, Inc., State College, USA).
3 Results and discussion
formed gels, which proves that the degree of hydrolysis reached
by the starch molecules in both species was not very extensive,
retaining the required properties, such as gel formation, for
starches to be used as fat substitutes (RADLEY, 1976).
NPS and NCS had the lowest gel texture characteristics
when compared to those of acid thinned. It is known that acid
treatment of starch causes partial hydrolysis of starch chains,
resulting in much lower paste viscosity. However, when the paste
cools down, acid-thinned starch chains tend to associate with
each other more easily, forming a more rigid gel (HOSENEY,
1994).
he ACS melted at 23 °C and APS, at 46 °C. Both showed gel
thermo-reversibility when subjected to cooling, ater melting.
he melting point of APS indicates similarity with fats that have
high melting point, which are more stable for storage under
high temperatures even though its use is not recommended as
a fat substitute in melt-in-the-mouth foods, such as ice cream
and chocolate. According to Giese (1996), fat replacing starches
work better in food systems with high humidity such as those
found in mayonnaise, salad dressings, and in meat emulsions.
In addition, they can be used in baked goods such as cakes, but
not in cookies due to their low moisture content.
3.3 Light transmittance (%)
he inluence of refrigerated storage on the paste clarity of
the starches is shown in Figure 1. he reduction in percentage
of transmittance of native and acid-thinned starches is a result
of retrogradation tendency (STAHL et al., 2007). he light
3.1 Dextrose equivalent measure
he DE value of APS (acid-thinned pinhão starch) was
6.5 ± 0.026. In the treatments with DE > 2.5 (percentage of
hydrolysis > 0.95), the gel thermo-reversibility can be observed,
which indicates a potential applicability of the starch as a fat
substitute since thermoreversible starch gel, similarly to fat,
has the ability to merge and solidify when exposed to high
and low temperatures, respectively. According to the National
Starch and Chemical Corporation (NATIONAL..., 1985), acidthinned starches with 5.0 ≤ DE < 8.0 form thermoreversible gels
of diferent consistencies, which are appropriate to be used as
fat replacers. According to Zambrano and Camargo (1999), fat
substitutes are obtained by starch hydrolysis when the degree of
hydrolysis (DE value) generated by the reaction is not excessive
(greater than 10) because when the acid hydrolysis overextends,
either by excessive conditions of time, temperature or acid
concentration, the gel-forming properties are lost and the starch
has no more use as a fat substitute.
Table 1. Formation, thermo-reversibility and texture characteristics
of native and acid-thinned starches from pinhão and corn varieties.
Variety
NPS
NCS
APS
ACS
Gel
formation
Yes
Yes
Yes
Yes
Gel
melting
No
No
Yes
Yes
Gel thermoreversibility
Yes
Yes
Gel texture
characteristics
sot
sot
hard
hard
NPS: Native pinhão starch, NCS: native corn starch, APS: acid-thinned pinhão starch,
ACS: acid-thinned corn starch
3.2 Formation, melting point, and thermo-reversibility of gel
starches
he properties of formation, melting point, and thermoreversibility of native and acid-thinned starches from pinhão
and corn varieties are shown in Table 1. he thermo-reversibility
analysis was performed in gels that melted under the conditions
evaluated. he native starches formed gels that did not melt ater
heating demonstrating that the acid modiication had a strong
efect on the properties of both starch species analyzed in this
study. he acid-thinned pinhão (APS) and corn starches (ACS)
Ciênc. Tecnol. Aliment., Campinas, 33(Supl. 1): 89-94, fev. 2013
Figure 1. Effect of refrigerated storage on the paste clarity (%
transmittance, at 625 nm) of native and acid-thinned starches from
pinhão and corn. he error bars represent the standard deviation
of mean. Native pinhão starch, ◊, native corn starch, □, APS, ∆ and
ACS, ○.
91
Evaluation of technological functional properties of acid-thinned pinhão starch
transmittance (%) of pastes of NPS and APS was higher than
that obtained for corn starches ater similar storage period,
which evidenced lower levels of retrogradation in pinhão
starches if compared to those of corn starches and, according
to Bello-Pérez et al. (2006), this is in agreement with the lower
content of amylose of pinhão starch (23.6%) in comparison to
that of corn starch (30%) and suggests the use of pinhão starch
in products stored for a long time and must have a sot texture,
such as some baked products.
APS and ACS pastes showed higher % transmittance in
comparison to those of NPS and NCS pastes. According to
Lawal (2004), the leaching of amorphous regions during acid
thinning enhances interactive bound formation between the
amylopectin molecules thus increasing its light transmittance.
his result agrees with that obtained by Sandhu, Singh and Lim
(2007), who found an efective increase in % transmittance in
the acid-thinned corn starch granules, when compared to that
of native corn starch.
3.4 Paste freezing and thaw stability
Figure 2 shows that NPS had a greater tolerance (syneresis
of 4.91%) than NCS (syneresis of 8.01%) ater 6 freeze-thaw
cycles. Stahl et al. (2007) reported water exudation value of
approximately 77% for native pinhão starch and about 82% for
native corn starch, both submitted to three cycles of freezing
(–18 °C) and thawing (30° C). he diference between these
results is probably due to differences in the measurement
method and the types of starch used in each experiment.
he lower syneresis of pinhão starch, when compared to
that of corn starch, makes it more suitable for use in custard,
puddings and pie-illings which are frozen stored (REGE; PAI,
1996; MARCON et al., 2007).
he acid-thinned starches had lower tolerance (extensive
syneresis) to the freeze-thaw cycles than the native starches. he
freeze-thaw stability of the acid-thinned starches was similar to
that found by Takizawa et al. (2004) and Shirai et al. (2007), who
attributed the high syneresis to a partial degradation of starch
macromolecular constituents during acid hydrolysis. According
to these authors, the fragmentation of starch chains during the
chemical treatment could be associated with the higher water
liberation due to intensive molecular re-association.
3.5 Swelling Power (SP) and solubility
Swelling power (SP) of NPS was strongly correlated to
the increases in temperature (Table 2). For native starches,
the increase in SP is related to the breaking of intermolecular
hydrogen bonds in amorphous areas allowing progressive water
absorption, as mentioned by Bello-Pérez et al. (2006). However,
for the native corn starch, the inluence of temperature on SP
was observed only above 70 °C , which, according to Li and
Yeh (2001), is due to the gelatinization of corn starch, which
starts only at 65.7 °C (T0, initial temperature of gelatinization),
as demonstrated by thermal analysis (DSC).
It can be said that acid-thinned starches difered from
native starches since their swelling power decreased with the
increase of temperature. his may be related to the fact that
during acid hydrolysis the amylose chains are fragmented
forming a disorganized structure that cannot retain water during
temperature increases (SANDHU; SINGH; LIM, 2007).
For the native species, at 25 and 60 °C, contrary to what
was reported by Wosiacki and Cereda (1989), pinhão starch
showed lower values of swelling power (p < 0.05) than those of
corn starch, while at 50 °C, no signiicant diference was found
between the species. However, at 70 °C, similarly to a study
reported by Stahl et al. (2007) and Bello-Pérez et al. (2006),
the value of the swelling power of pinhão starch was higher
compared to that of corn starch. According to these authors, this
inding is expected from the known inverse correlation between
swelling power and amylose and lipid levels at temperatures
close to 85 °C.
he SP of APS and ACS was higher (p < 0.005) than that
of similar native starches at temperatures of 25 °C and 50 °C,
whereas at 60 °C and 70 °C, the opposite was observed, as
similarly reported by Sandhu, Singh and Lim (2007) when
assessing acid-thinned corn starch. According to Lawal (2004),
this is due to the reduction of the amorphous region of the
starch granule, which reduces the number of sites that establish
links with the water molecule. Studies have shown that genetic
variations, climate and soil conditions, maturation stage, and
Table 2. Swelling power of native and acid-thinned starches (pinhão
and corn).
Temperature
25 °C
50 °C
60 °C
70 °C
Figure 2. Effect of freeze-thaw cycles on the syneresis (% water
separated ater 6 cycles) of native and acid-thinned starches from
pinhão and corn varieties.
92
Swelling power (g water/g starch)
NPS
NCS
APS
ACS
1.76aA ± 0.13 1.93bA ± 0.01 2.16cA ± 0.01 2.34dA ± 0.00
1.91aB ± 0.01 1.92aA ± 0.00 2.25bB ± 0.02 2.27bB ± 0.02
3.84aC ± 0.02 4.18bB ± 0.01 2.07cC ± 0.03 2.18dC ± 0.01
11.08aD ± 0.04 4.82bB ± 0.02 1.55cD ± 0.00 1.66dD ± 0.00
he results are expressed as mean ± standard deviation (n = 3). Means followed by
diferent lowercase letters in the same row indicate signiicant diferences by Tukey
test (p < 0.05). Means followed by diferent capital letters in the same column indicate
signiicant diferences by Tukey test (p < 0.05).
Ciênc. Tecnol. Aliment., Campinas, 33(Supl. 1): 89-94, fev. 2013
Thys et al.
Table 3. Solubility of native and acid-thinned starches from pinhão
and corn.
Temperature
25 °C
50 °C
60 °C
70 °C
NPS
0.31aA ± 0.04
0.12aB ± 0.01
4.02aC ± 0.20
1.45aD ± 0.09
Variety
NCS
APS
0.71bA ± 0.01 7.52cA ± 0.12
1.48bB ± 0.04 7.47cA ± 0.12
2.98bC ± 0.11 8.65cB ± 0.02
2.85bC ± 0.06 9.02cC ± 0.05
ACS
6.87dA ± 0.07
6.93dA ± 0.14
8.11dB ± 0.04
8.19dB ± 0.02
he results are expressed as mean ± standard deviation (n = 3). Means followed by
diferent lowercase letters in the same row indicate signiicant diferences by Tukey
test (p < 0.05). Means followed by diferent capital letters in the same column indicate
signiicant diferences by Tukey test (p < 0.05).
harvest time also afect the swelling power (FRANCO et al.,
2002), which could explain the irst situation mentioned above.
Man et al. (2012) reported that no signiicant diference between
native and acid-thinned starch was found under 65 °C, but, at
temperatures higher than 80 °C, the SP gradually decreased.
he solubility (Table 3) of NPS was lower than that of
NCS at 25, 50, and 70 °C, which can be explained by the lower
amylose content of pinhão starch compared to that of corn
starch. Amylose dissociates from the granule, which contributes
to solubility increase (MARCON et al., 2007). Wosiacki and
Cereda (1989) reported a similar pattern for pinhão starch
at temperatures higher than 85 °C. Bello-Pérez et al. (2006)
reported an inverse behavior, with higher solubility values for
pinhão starch, which according to these authors is in agreement
with the lower temperature and enthalpy of gelatinization of
pinhão starch assessed by DSC in their study.
APS and ACS showed higher values of solubility than
the native starches. his occurs because the acid hydrolizes
preferentially the amorphous region of the starch molecule , where
amylose is normally found (ATICHOKUDOMCHAI et al.,
2000), generating a signiicant reduction in the amylose chain
length in the granule content, and its consequent dissolution
resulting in solubility increase; fact that was evidenced with
increases in temperature in the corn and pinhão species.
In addition, APS had significantly higher (p < 0.05)
solubility than ACS showing a higher susceptibility of pinhão
starch to acid hydrolysis, when compared to that of corn starch.
4 Conclusion
The functional properties of NPS (lower levels of
retrogradation and syneresis and highest solubility when
compared to native corn starch), which is a non-conventional
source of starch, suggest it may have potential use in food
systems. he acid hydrolysis (3.2 mol.L–1 HCl and 44 °C) of
pinhão and corn starches caused gel thermo-reversibility,
lower tendency to retrogradation of starch pastes, and higher
solubility at an economically viable reaction time (6 hours).
Furthermore, the APS showed a melting point (46 °C) close to
that of the conventional fats (37-45 °C), which may indicate that
the pinhão starch could be used as a fat substitute when the gel
is prepared (5%, w/w dry basis, total weight 28 g) by heating at
95 °C for 30 minutes. However, this applicability is limited to
Ciênc. Tecnol. Aliment., Campinas, 33(Supl. 1): 89-94, fev. 2013
frozen or refrigerated food since the acid hydrolysis reduced
the tolerance of both starches to refrigerate storage (5 °C) and
to the freeze-thaw cycles.
Acknowledgements
he authors thank Florencia Cladera-Olivera and Mauricio
Seibel Luce for the critical reading of the manuscript and very
helpful discussions and comments.
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