Corresponding author. Tel.: +54 739 42020; fax: +52 739 41896.
E-mail address: labellop@ipn.mx (L.A. Bello-Pe rez).
(Drzikova, Dongowsky, Gebhardt, & Habel, 2005). In the
digestive tract, DF exerts a buffering effect that links excess
of acid in the stomach, increases the fecal bulk and
stimulates the intestinal evacuation; besides, it provides a
favorable environment for the growth of the benecial
intestinal ora. Fibre can also bind diverse substances
including cholesterol (Jenkins, Kendall, & Ransom, 1998;
Jime nez-Escrig & Sa nchez-Muniz, 2000). It has been
reported that these specic properties of DF play an
important role in the prevention and treatment of obesity,
atherosclerosis, coronary heart diseases, colorectal cancer
and diabetes (Bingham, 2003; Peters, 2003; Terry, 2001).
Consumption of certain bres, particularly highly viscous
SDFs, is usually associated with moderate postprandial
glycemic responses, a property of importance in the dietetic
treatment of diabetes (Jenkins et al., 1998; Schneeman,
1987).
There is a trend to nd new sources of DF as ingredients
for the food industry (Chau & Huang, 2003). The most
widespread consumed DF products are those derived from
cereals. However, over the past decade high DF materials
from fruits (citrus, apple, and others) have been steadily
introduced in the occidental world markets. Fruit DF
concentrates have, in general, better nutritional quality
than those found in cereals, because of their signicant
contents of associated bioactive compounds (avonoids,
carotenoids, etc.) and more balanced composition (higher
overall bre content, greater SDF/IDF ratio, water- and
fat-holding capacities, lower metabolic energy value, and
phytic acid content) (Chau & Huang, 2003; Jime nez-Escrig,
Rinco n, Pulido, & Saura-Calixto, 2001).
According to recent reports (Saura-Calixto, 1998), the
presence of signicant amounts of bioactive compounds,
such as avonoids and carotenoids, in DF from fruits
imparts them considerable nutritional value. The antiox-
idant properties of avonoids and carotenoids come from
their ability to link free radicals that easily attack saturated
fatty acids present in cell membranes, causing peroxida-
tion, decreasing permeation and damage of membrane
proteins, leading to cellular inactivation. DNA is also
subject to free radical-effects producing mutations which
may lead to cancer (Ubando, Navarro & Valdivia, 2005;
Yen & Hsieh, 1998).
Mango is a climacteric fruit that in Me xico and many
other countries is consumed in ripe state. For this
reason, large quantities of fruit are lost during commercia-
lization, as consequence of decient postharvest handling.
Unripe mango fruit represents an alternative source
for DF production, due to a number of reasons such
as, starch content of the pulp, high cellulose, hemi-
cellulose, lignin and carotenoid levels, as well as the low
cost of the fruit that may allow the preparation of DF
concentrates with attractive chemical and functional
characteristics.
The aim of the present work was to evaluate the chemical
and functional properties of DF from unripe mango fruit
and its application in a bakery product.
2. Materials and methods
2.1. Sample preparation
Commercial hard green preclimacteric mango (Mangi-
fera indica L. var. Tommy Atkins) fruits were purchased
from the local market of Cuautla, Morelos, Me xico. They
were washed and cut into 1 cm slices in order to obtain the
fruit pulp with the skin. The slices was treated as described
by Larrauri, Rupe rez, Borroto, and Saura-Calixto (1996)
for the isolation of the brous fraction (mango dietary
bre, MDF), which was dried at 50 1C and kept at room
temperature in sealed plastic containers for further
analyses.
2.2. Chemical composition
Moisture content was determined by gravimetric heating
(13072 1C for 2 h) using a 23 g sample. Ash, protein and
fat were analysed according to AACC methods 0801,
4613 and 3025, respectively (AACC, 2000). DF was
tested using the 985.29 AOAC method (AOAC, 1999). DF
fractions were obtained as indigestible residues after
enzymatic digestion of non-DF components; the insoluble
residues were isolated by ltration, and soluble bre was
precipitated with ethanol. Dried residues correspond to
insoluble dietary bre (IDF) and soluble dietary bre
(SDF), respectively. Total starch (TS) was determined by
the method of Gon i, Garc a-Alonso, and Saura-Calixto
(1997); in brief, 50 mg of sample were dispersed in 2 M
KOH (30 min) to hydrolyze all the starch, then samples
were incubated with amyloglucosidase (Boehringer, No.
102857, 60 1C, 45 min, pH 4.75), and glucose was deter-
mined using the glucose oxidase assay GOD-POD. TS was
calculated as glucose (mg) 0.9; potato starch was used as
a control. Total soluble carbohydrates were assessed by the
phenolsulphuric method (Dubois, Gilles, Hamilton, Re-
bers, & Smith, 1956).
2.3. Extractable polyphenols (EPPs)
EPPs were extracted from samples using aqueousor-
ganic solvents. The extraction procedure is described
elsewhere (Jime nez-Escrig, Jime nez-Jime nez, Pulido, &
Saura-Calixto, 2001). The supernatants were combined
and total extractable phenols were estimated by the
FolinCiocalteau method (Jime nez-Escrig et al., 2001)
using gallic acid as a standard; results were expressed as
gallic acid equivalent (GAE).
2.4. Antioxidant activity (DPPH free-radical scavenging)
assay
Supernatants containing EPP obtained as described
above were used to determine the antioxidant capacity of
mango dietary bre. It was measured in terms of radical
scavenging ability, according to the DPPH
d
method
ARTICLE IN PRESS
N. Vergara-Valencia et al. / LWT 40 (2007) 722729 723
(Sanchez-Moreno, Larrauri, & Saura-Calixto, 1998). The
parameter EC
50,
which reects 50% depletion of DPPH
d
free-radical, was expressed in terms of grams of mango
dietary bre equivalent per gram of DPPH
d
in the reaction
medium. The time necessary to reach the steady state at
EC
50
(T
EC50
) was calculated. The antiradical efciency
(AE), an index dened by Sanchez-Moreno et al. (1998)
which combines both factors, was also calculated in order
to recognize the combined effect of both parameters:
AE 1/EC
50
T
EC50
.
2.5. Water (WHC) and oil (OHC) holding capacities
Twenty-ve milliliters of distilled water or commercial
olive oil were added to 250 mg of dry sample, stirred and
left at 40, 60 or 80 1C for 1 h. After centrifugation, the
residue was weighed and WHC and OHC calculated as g
water or oil per g of dry sample, respectively (Larrauri
et al., 1996).
2.6. Preparation of bakery products
Raw materials were acquired in local supermarkets
(Yautepec, Mexico) and stored in glass or plastic contain-
ers at room temperature (25 1C) or under refrigeration,
depending on the recommended storage conditions of each
material. Formulations for the bakery products are shown
in Table 1. Basically, the experimental preparations
contained MDF in exchange for commercial wheat germ
(Tres Estrellas, Estado de Me xico, Me xico). The dough for
the products was prepared by mixing of the ingredients; in
the case of cookies, the dough was extended immediately
until obtaining a surface of 2 cm height, and cookies were
cut with a circular mould (6 cm diameter), and placed on
greased aluminium cookie sheets. Bread dough was
fermented during 30 min at room temperature and then
transferred into greased aluminium moulds. Both types of
items were baked in a household oven (Hotpoint,
6B4411LO, Leisser S.A. de C.V., San Luis Potos , Me xico),
at an approximate temperature of 200 1C for 20 min. Once
baked, the products were allowed to cool down during
30 min and then stored in plastic containers with hermetic
cover. Control products, based on wheat germ, were
chosen as a pragmatic approach.
2.7. Experimental studies in bakery products
Chemical composition, EPPs, Antioxidant activity and
DPPH free-radical scavenging assays were carried out as
above.
2.8. Available starch
Available starch content in bakery products was
measured with the multienzymatic method described by
Holm, Bjorck, Drew, and Asp (1986).
2.9. Starch hydrolysis index of products as eaten
(chewing/dialysis test)
The in vitro rate of starch hydrolysis in bakery products
was assessed with the protocol developed by Granfeldt,
Bjorck, Drews, and Tovar (1992). Samples of baked
products, containing 1 g of available starch, were tested.
Six healthy subjects participated in the chewing phase of
the experiments, which was followed by pepsin digestion
and further incubation with porcine pancreatic amylase in
a dialysis bag. The reducing amylolysis products appearing
in the dialysate were measured colorimetrically, and
expressed as maltose equivalents. Data were plotted as
hydrolysis degree versus time curves and the hydrolysis
index (HI) was calculated as the area under the curve
(0180 min) for the test product expressed as a percentage
of the corresponding area for commercial decorticated
white bread (used as a reference), chewed by the same
person. The average HI was calculated from the six
digestion replicates run for each sample and means were
compared by the MannWhitney test, using a statistical
program (version 2.03, SPSS, Chicago, IL). The predicted
glycemic index (pGI) was calculated from HI values, using
the empirical formula proposed by Granfeldt (1992)
pGI 0.862 HI+8.198, for which the correlation coef-
cient (r) is 0.96 (Po0:001).
2.10. Statistical analysis
Results were expressed as mean values7standard error
of at least two separate determinations. Comparison of
means was performed by one-way analysis of variance
(ANOVA) followed by Tukeys test. Statistical analyses
were run using the computer SPSS V. 6.0 software (SPSS
Institute Inc., Cary NC).
ARTICLE IN PRESS
Table 1
Formulation of bakery products using mango dietary bre (MDF)
Product type
Ingredient (g) Cookie
with MDF
Control
cookie
Bread with
MDF
Control
bread
Wheat our 25 25 60 60
MDF 75 40
Regular margarine 25 25 25 25
Onion 1.5 1.5 1.5 1.5
Garlic 1.5 1.5 1.5 1.5
Pepper 0.5 0.5 0.5 0.5
Yeast 5 5 5 5
Salt 2.5 2.5 2.5 2.5
Sugar 3.0 3.0 3.0 3.0
Milk 80 80 80 80
Wheat Germ 75 40
N. Vergara-Valencia et al. / LWT 40 (2007) 722729 724
3. Results and discussion
3.1. Chemical composition
MDF exhibited low moisture content (Table 2), similar
to those determined in commercial dry products such as
wheat germ, oat cookies and bran akes (Sangronis &
Rebolledo, 1993). An important parameter in a bre
concentrate is its lipid content; MDF had a lipid content of
2.3 g/100 g dry sample, lower than those reported in grape
skins (6.877.78 g/100 g dry sample) (Bravo & Saura-
Calixto, 1998), commercial grape bre (6.9 g/100 g dry
sample) (Saura-Calixto, 1998) and the peel of Citrus
sinensis (22.2 g/100 g dry sample) (Chau & Huang, 2003).
MDF might be therefore considered a potential ingredient
for the formulation of light food products. Protein content
in MDF was also low (4.2 g/100 g dry sample) as compared
to other bres such as grape skins (11.614.4 g/100 g dry
sample) (Bravo & Saura-Calixto, 1998) and a commercial
bre (14.4 g/100 g dry sample) obtained from red grape
peels (Saura-Calixto, 1998). However, a lower protein
value (2.2 g/100 g dry sample) was determined in the bre
concentrate from Citrus sinensis peel (Chau & Huang,
2003). Fruits are characterized by their content of certain
minerals components; in MDF a 2.8 g/100 g dry sample ash
content was recorded, which is similar to that determined
(3.3 g/100 g dry sample) in bre from Citrus sinensis peel
(Chau & Huang 2003), but lower than the in grape skins
(between 5.7 and 9.2 g/100 g dry sample) (Bravo & Saura-
Calixto, 1998; Saura-Calixto, 1998).
It is important to mention that the MDF preparation
was obtained from the whole fruit (pulp and peel). Pulp of
unripe mango is characterized by containing starch (Bello-
Pe rez, Ottenhof, Agama-Acevedo, & Farhat, 2005; Kaur,
Singh, Singh-Sandhu, & Singh-Guraya, 2004). TS content
in the bre concentrate was of 29.8 g/100 g dry sample
(Table 2); such a high starch level that could contribute to
the formation of resistant starch (RS) during processing of
MDF-based products, as it has been shown in mango
starch extrudates (Agustiniano-Osornio et al., 2005). Total
dietary bre (TDF) content in MDF was 28.1 g/100 g dry
sample (Table 2), which is close to levels measured in
antioxidant DF preparations from guava (4849 g/100 g
dry sample) (Jime nez-Escrig, Jime nez-Jime nez et al., 2001;
Jime nez-Escrig, Rinco n, Pulido, & Saura-Calixto, 2001),
although notably lower than TDF contents reported in
grape skins (54.164.6 g/100 g dry sample) (Bravo & Saura-
Calixto, 1998; Saura-Calixto, 1998), citrus peel (57 g/100 g
dry sample) (Chau & Huang, 2003), in bre from two
Mexican lime peels (66.7 and 70.4 g/100 g dry sample)
(Ubando et al., 2005) and in mango peel bre (6571 g/
100 g dry sample) (Larrauri et al., 1996). The relatively low
TDF content in MDF concentrate relates to the high starch
content. However, for certain kind of food products such a
starch level might be of importance, given the additional
functional properties imparted by starch. For certain food
applications, DF concentrates should have a balanced
content of soluble (SDF) and insoluble (IDF) fractions; in
the case of MDF the two bre types are present at almost
equal levels. Thus, it exhibits a good balance that might be
also important from the nutritional point of view. DF of
Mexican lime peel contains a higher proportion of IDF
(Ubando et al., 2005); a similar pattern has been reported
for other bre preparations, such as citrus peel bre, where
47.6 g/100 g dry sample IDF and 9.4 g/100 g dry sample
SDF were determined (Chau & Huang, 2003), guava bre
(39.250.1 g/100 g dry sample IDF and 1.5 and 1.8 g/100 g
dry sample SDF) (Jime nez-Escrig, Jime nez-Jime nez et al.,
2001; Jime nez-Escrig, Rinco n et al., 2001) and mango peel
(43.4 g/100 g dry sample IDF and 24.7 g/100 g dry sample
SDF) (Larrauri et al., 1996). Total soluble carbohydrates in
MDF accounted for 32.6 g/100 g dry sample; most of this
value must correspond to simple sugars. This level is much
higher than those reported in grape skins (3.0 g/100 g dry
sample) (Bravo & Saura-Calixto, 1998; Saura-Calixto,
1998), but not so different from those reported in guava
bre (28 g/100 g dry sample), citrus peel bre (27.3 g/100 g
dry sample) (Chau & Huang, 2003) and bre from mango
peel (26.7 g/100 g dry sample) (Larrauri et al., 1996).
Extractable polyphenols or total soluble polyphenols
(EPP) content in MDF was 16.1 mg/g, which is higher than
in apple bre (3 mg/g) (Larrauri et al., 1996), but similar to
those reported in bre from Mexican lime peels (10.55 and
19.9 mg/g) (Ubando et al., 2005) and in a commercial grape
skin bre (20 mg/g) (Saura-Calixto, 1998). However, EPP
in MDF was lower than those determined in two different
antioxidant guava bres (26.3 and 58.7 mg/g) (Jime nez-
Escrig, Jime nez-Jime nez et al., 2001; Jime nez-Escrig,
Rinco n et al., 2001), grape skins (between 37.6 and
52.2 mg/g) (Bravo & Saura-Calixto, 1998) and in bre
from mango peel (between 44 and 70 mg/g) (Larrauri et al.,
1996). EPP are low or intermediate molecular mass
phenolics that are extracted using different solvents and
appear to be absorbed in the digestive tract, exerting
systemic effects (Saura-Calixto, 1998). The anti-radical
efciency (AE) was tested in EPP obtained from MDF,
ARTICLE IN PRESS
Table 2
Chemical composition (g/100 g dry sample, except for moisture),
extractable polyphenols (mg/g dry sample) and anti-radical efciency
(10
3
) of mango dietary bre
Component Amount
Moisture 8.1070.02
Lipid 2.3570.02
Protein 4.2870.11
Ash 2.8370.01
Total starch 29.8870.06
Insoluble dietary bre 13.8070.71
Soluble dietary bre 14.2570.71
Total dietary bre 28.0570.24
Total soluble carbohydrates 32.6270.25
Extractable polyphenols 16.1470.24
Anti-radical efciency (AE) 15.0370.07
Mean of three replicates 7 standard error.
N. Vergara-Valencia et al. / LWT 40 (2007) 722729 725
obtaining a 15 10
3
value; this was higher than those
determined in bres extracted from guava fruit
(7 10
3
9 10
3
), which were considered suitable for
antioxidant DF production (Jime nez-Escrig, Jime nez-
Jime nez et al., 2001; Jime nez-Escrig, Rinco n et al., 2001);
hence, present results point out MDF as an important
antioxidant DF source.
3.2. Functional properties of MDF
The results of WHC and OHC are presented in Fig. 1.
MDF showed increased WHC values with temperature,
although no statistical differences were found at the highest
temperature assessed. This behaviour might be due to the
high SDF (Agustiniano-Osornio et al., 2005) and starch
contents present in this bre preparation. The value
determined at low temperature agrees with that reported
in mango peel DF (11 g of water/g of dry sample) (Larrauri
et al., 1996). Other WHC reported are in the range of
15.516.7 g of water/g of dry citrus peel bre (Chau &
Huang, 2003) and between 6.96 and 12.84 g of water/g of
dry Mexican lime peel bre. OHC is another important
parameter, which in MDF ranged between 1.0 and 1.5 g of
oil/g of dry sample. These values were lower than those
reported in other bre concentrates, e.g. mango peel DF
(4 g of oil/g of dry sample) (Larrauri et al., 1996) and in
citrus peel bre (2.355.09 g of oil/g of dry sample) (Chau
& Huang, 2003). Accordingly, MDF the use of bre may
be appropriate in products where emulsifying properties
are not required.
3.3. Chemical composition of bakery products
Moisture content was similar in control and MDF-
containing cookies (Table 3), showing that moisture was
not affected by the indigestible fraction added in the
formulation. However, MDF-containing bread was drier
than the control sample (a 0:05), indicating that in this
kind of product formulation may affect this important
parameter, critical for its stability and shelf-life (Bushuk,
1998).
Only minor differences between control and MDF-
containing samples were found regarding fat and protein
contents (Table 3). The higher level of both components in
control products must be due to the inclusion of wheat
germ, an ingredient providing signicant amounts of lipids
and proteins (Atwell, 2001). Ash content did not vary
between control and experimental products nor did TS,
indicating that wheat germ and MDF made a similar
contribution of these components to the nal products.
Since less MDF was used in the bread than in the
experimental cookie, the later had lower TS content.
ARTICLE IN PRESS
0.0
0.2
0.4
0.6
0.8
1.0
1.2
1.4
1.6
1.8
40 60 80
Temperature (C)
O
H
C
(
g
o
i
l
/
g
d
r
y
s
a
m
p
l
e
)
0
(a)
(b)
5
10
15
20
40 60 80
Temperature (C)
W
H
C
(
g
w
a
t
e
r
/
g
d
r
y
s
a
m
p
l
e
)
Fig. 1. Water-holding capacity (WHC) (a) and oil-holding capacity
(OHC) (b) of mango dietary bre.
Table 3
Chemical composition (g/100 g dry sample, except for moisture), total extractable phenol content (mg/g dry sample) and anti-radical efciency ( 10
3
) of
bakery products added with mango dietary bre (MDF)
Bakery product
Component Cookie with MDF Control cookie Bread with MDF Control bread
Moisture 11.970.0
a
12.070.1
a
26.070.1
b
31.570.9
c
Fat 13.070.3
a
13.870.1
b
11.270.1
c
16.270.2
d
Prot 8.370.0
a
9.170.1
b
10.270.2
c
11.470.1
d
Ash 4.370.0
a
4.370.0
a
3.970.0
b
3.270.0
c
Total starch 45.570.8
a
48.970.9
b
48.470.7
b
67.270.4
c
Insoluble dietary bre 11.470.4
a
12.070.3
a
9.470.5
b
4.370.5
c
Soluble dietary bre 6.070.4
a
1.370.3
b
7.170.5
c
6.070.4
a
Total dietary bre 17.470.1
a
13.370.1
b
16.670.1
c
14.270.1
d
Total extractable polyphenols 11.871.2
a
nd 10.170.5
a
nd
Anti-radical efciency (AE) 9.170.6
a
nd 8.670.3
a
nd
Available starch 41.170.2
a
44.170.1
b
41.170.5
a
60.971.0
c
Means in row not sharing the same superscript letter are signicantly different (Po0:05) (means of three replicates), nd no detected.
N. Vergara-Valencia et al. / LWT 40 (2007) 722729 726
Both products formulated with MDF exhibited in-
creased TDF levels (Table 3), with a better balance of
SDF and IDF. In MDF-bread the IDF increase was more
than 100 g/100 g dry sample. Addition of MDF improves
the SDF content of cookies signicantly, as the control
sample presented only 1.3 g/100 g dry sample SDF whereas
the MDF-containing product exhibited a 6.0 g/100 g dry
sample level. TDF contents in commercial bre-rich
cookies have been reported between 3.73 and 5.95 g/100 g
dry sample with marked predominance of IDF (Sangronis
& Rebolledo, 1993), thus resembling the pattern presented
here. Commercial integral bread samples, on the other
hand, had TDF contents of 4.996.11 g/100 g dry sample,
with predominance of IDF (Sangronis & Rebolledo, 1993).
Bakery products prepared here with MDF concentrate had
interesting TDF contents and may be an alternative for
people with special caloric requirements. Process-induced
changes in the SDF/IDF ratio in baked goods containing
MDF deserve further investigation.
Both types of bakery products containing MDF
exhibited similar levels of EPPs, although they were slightly
lower than in the original MDF concentrate (Tables 2 and
3). These results suggest that baking does not have
signicant impact on these compounds. In the same sense,
despite the fact that the baked-products showed apparently
decreased AE of values, they were higher than those
determined in bre from guava fruit (Jime nez-Escrig,
Jime nez-Jime nez et al., 2001; Jime nez-Escrig, Rinco n,
Pulido et al., 2001), stressing that products baked with
MDF have signicant antioxidant properties.
3.4. Starch bioavailability
Bakery products added with MDF presented similar
mango dietary bre concentrate (AS) content (Table 3).
Absolute AS values in the MDF goods appear lower than
in control samples. However, when AS is calculated as a
percentage of TS in the cookies, such a difference is ruled
out indicating that the variation is mainly due to starch
dilution created by the use of wheat germ, a material of low
starch content. A larger difference in AS was evident
between the two breads, since AS content in control is
approximately 50 g/100 g dry sample more than that of the
MDF-containing product. Thus, differential formation of
resistant starch (RS) fractions upon baking (Bello-Pe rez,
Sa yago-Ayerdi, Me ndez-Montealvo, & Tovar, 2004; Gran-
feldt, 1994; Liljeberg, A