functional foods, antioxiadant

1. Ph ton 227
The Journal of Food Technology. Photon 106 (2014) 227-238
https://sites.google.com/site/photonfoundationorganization/home/the-journal-of-food-technology
Original Research Article. ISJN: 3852-1875: Impact Index: 3.43
The Journal of Food Technology Ph ton
Evaluation of the addition of dry soybean sprouts on cooking yield
and oxidative stability of enriched beef patties with soybeans oil
Romero Mara Cristinaa,b*
, Garro Oscara,b
, Romero Ana Maríaa
, Doval Mirtha Marinaa
, Judis María
Aliciaa
a
Laboratorio de Industrias Alimentarias II, Universidad Nacional del Chaco Austral, Cte. Fernández 755, (3700)
P.R. Sáenz Peña, Chaco, Argentina
b
CONICET, Consejo Nacional de Investigaciones Científicas y Técnicas, Argentina
The authors receive Thomas Edison Award-2014 in
Food Technology for Inspiration and Knowledge
Distribution among young research scholars.
Article history:
Received: 29 January, 2014
Accepted: 02 February, 2014
Available online: 15 March, 2014
Keywords:
Beef patties characteristics - healthier lipids - lipid
oxidation - natural antioxidants - Glycine max L
Corresponding Author:
Romero M.C.
Professor
Email: mara@uncaus.edu.ar
Phone: 543644420137
Garro O.
Professor
Email: garro@uncaus.edu.ar
Phone: 543644420137
Romero A.M.
Professor
Email: amr@uncaus.edu.ar
Phone: 543644420137
Doval M.M.
Professor
Email: mdoval@uncaus.edu.ar
Phone: 543644420137
Judis M.A
Professor
Email: judis@uncaus.edu.ar
Phone: 543644420137
Abstract
The improvement of fatty acid profile of beef patties
through the replacement of pork back fat with
soybean oil can be achieved, but it is necessary to
use antioxidants, such as dry soybean sprouts, to
maintain the oxidative stability. Besides,
development of functional foods with soybean
sprouts as food ingredient opens up new
possibilities to their use as natural additive. This
work was designed to investigate the effect of the
addition of soybean sprouts on the quality
characteristics of beef patties with partial
replacement of pork back fat by soybean oil during
frozen storage (90 days at –18 ± 2 ºC). Results
showed that addition of soybean sprouts improved
protein and carbohydrate content; moisture and oil
retention of cooked enriched patties were also
enhanced. One percent (1%) of dry soybean
sprouts (DSS) concentration was the most effective
to retain healthier lipids and minimize the cooking
loss. Besides, these products were microbiological
stable and had a softer texture than those without
soybean sprouts addition. This study indicates that
proximate composition could be improved through
incorporation of dry soybean sprouts, and it could
be used to protect enriched beef patties from lipid
oxidation without modifying their quality
characteristics.
Citation:
Romero M.C., Garro O., Romero A.M., Doval M.M., Judis
M.A., 2014. Evaluation of the addition of dry soybean
sprouts on cooking yield and oxidative stability of enriched
beef patties with soybeans oil. The Journal of Food
Technology. Photon 106, 227-238.
All Rights Reserved with Photon.
Photon Ignitor: ISJN38521875D654215032014
1. Introduction
In recent years, the demand for fast food has
been increasing rapidly. Since it is generally
recognized that meat and meat products make
an important contribution to nutrition,
numerous efforts have been made to optimize
their composition in order to help consumers
adapt their diet to nutrient intake goals.
Burgers and patties are common meat
products widely accepted by all population,
and the possibility of inducing changes of
composition to improve their nutritional value
and their potential health-beneficial properties
is among the advantages they present (López-
López et al., 2010). Various ground beef
products have been developed by replacing

2. Ph ton 228
animal fat with vegetable oil so as to be more
in line with health recommendations by
reducing saturated fatty acids (SFA) and
cholesterol, and increasing monounsaturated
fatty acids (MUFA) and polyunsaturated fatty
acids (PUFA) (Dzudie et al., 2004; Hur et al.,
2008; Lee et al., 2005). Therefore, in order to
development functional meat products, liquid
olive oil has been added to beef patties (Hur et
al., 2008), also the addition of pre-emulsified
soybean oil as replacement of pork back fat
was studied (Muguerza et al., 2003); but this
addition decrease the cooking yield and affect
the oxidative stability of the enriched product.
Soy proteins are widely used in meat products
for their nutritional value and functional
properties in the form of flours, concentrates
and isolates, since they increase water
retention, fat retention, emulsion stability,
nutritional content and cooking yield. Several
researchers (Porcella et al., 2001; Chin et al.,
2000; Feng et al., 2003) informed that soy
protein isolates (SPI) can be used as additives
in foods, especially in meat products,
improving functional properties of the system
such as water binding and textural properties.
A further advantage of soy proteins is the
antioxidant activity of some fractions of them
(Peña-Ramos and Xiong, 2003; Romijn et al.,
1991).
Germinated soybean sprouts are
commercialized in our country as raw staple
vegetables and used in soups and salads. It is
particularly promising as meat ingredient due
to its antioxidant compounds as vitamin C 475
mg/kg dry matter, phenolics compounds 391
mg/g dry sprouts, flavonoids content 184 mg/g
quercetin equivalents of dry sprouts and
superoxide dismutase (SOD) activity in crude
extracts from dried soybean sprouts (DSS)
was 3110 unit/g dry matters. In previous
research, the effectiveness of different
concentrations of DSS as antioxidant on
cooked chicken patties stored at 6 ± 1° C for 8
days was evaluated, showing a decrease of
lipid oxidation in all cases, although they were
only strongly effective to concentrations higher
than 30 g/kg (Romero et al., 2008).
Even when soybean oil has been added to
pork patties in liquid form (Jung and Joo,
2013) and the DSS have been investigated
previously (Romero et al., 2008), there are no
reports with the effect these products on the
chemical composition and- or on technological
properties during frozen storage of functional
cooked meat products.
2. Objective of Research
Consequently, this work was designed (1) to
evaluate the effects of the pork fat partial
replacement by soybean oil and the addition
dried soybean sprouts on the proximate
composition and cooking properties of beef
patties and (2) to investigate the effectiveness
of DSS on reducing lipid oxidation of enriched
cooked beef patties during frozen storage (90
days at –18 ± 2° C). Also, texture and
microbiology analyses were carried out over
the samples that showed higher oil retention
and acceptable oxidative stability
3. Materials and Methods
3.1 Study area
The soybean (Glycine max L.) was obtained
from soybean cultivars in the province of
Chaco situated between the parallels 24° and
28° south latitude and between the meridians
58° and 63° west longitude, Argentina
(America). This work was carried out between
March of 2012 and July of 2013. This enriched
beef patties with DSS and soybean oil
provides both nutritional and phytochemical
benefits of soybean to human population.
3.2 Materials
Dry soybean sprouts were obtained from
previously selected soybeans (Glycine max L.
Merr, Nidera A8009RG, Chaco, Argentina)
using as selection criteria hygienic condition,
shape and size. The beans were soaked in
water for one hour at 25° C and then
germinated in darkness at 30° C in a
controlled temperature chamber. Once the
sprouts reached 3 cm of length approximately,
they were separated from the beans and
dehydrated at 30° C in a static drying chamber
until 89% dry weight. Then dehydrated
soybean sprouts were ground to a powder.
3.3 Patties preparation
Enriched beef patties were prepared using
fresh lean beef meat and pork back fat
obtained from local markets, and soy oil
containing SFA 16 g/100g, MUFA 35 g/100g,
PUFA n6 60 g/100g and n3 6 g/100g (Aceitera
General Deheza S.A., Córdoba, Argentina)
that was used as fat partial replacer.
Six different formulations were elaborated as
follows: one control sample without addition of
antioxidants (C) with beef and 20% of pork
back fat (similar to commercial products); and
four enriched beef patties (EP) with beef, 10%
of pork back fat, 10% of soybean oil and with
addition of DSS as natural antioxidant (0%,

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0.5%, 1% and 2%), and 0.01% of butylated
hydroxyanisole (BHA) as antioxidative positive
control.
All formulations were shaped using Petri-dish
(90 mm x 20 mm) to obtain 60 patties of
approximately 100g each one. Half of the
samples were cooked in an electric oven (200°
C) until internal temperature reached 72° C.
Cooking properties, proximate analysis, fatty
acid composition, and texture profile were
evaluated within 8 h after the samples were
prepared.
To evaluate oxidative stability, the cooked
samples were packaged in oxygen permeable
bags (2000 cm
3
/m
2
day) using a packing
machine (RAPI-VAC S-750® SERVIVAC
S.R.L., Buenos Aires. Argentina) stored at -18°
C and analyzed at 0, 15, 30, 60 and 90 days.
All chemical and physical analyses were
carried out in duplicated for each formulation
and heat treatment (raw or cooked).
3.4 Proximate composition
Moisture, protein, fat and ash contents were
analyzed according to AOAC (1999) methods.
Total carbohydrates were quantified using
anthrone Clegg’s methods (1956) on an UV-
Vis Beckman DU 640B® (Fullerton, CA, USA)
spectrophotometer. Sodium content was
determined by a JENWAY PFP7® flame
photometer (Burlington, NJ, USA) in pre-ashed
and HCl acidified samples.
3.5 Determination of cooking properties
Percent cooking yield was determined by
calculating weight differences for samples
before and after cooking as follows (Naveena,
et al., 2006):
Fat retention percentage was calculated using
the following equation (El-Magoli et al., 1996):
Moisture retention represents the percentage
of moisture retained in the cooked product and
was determined according to Alakali, et al.
(2010):
Diameter and thickness of patties were
measured with a Vernier caliper. Shrinkage
percentagewas determined using the
equations reported by Serdaroǧlu (2006):
3.6 Fatty acids composition
Total lipids were extracted by Bligh and Dyer
(1959) method. Boron trifluoride/methanol was
used for the preparation of fatty acid methyl
esters (AOAC 1999) and then they were
analyzed using GC Mass Spectrometer
(Thermo Fisher Scientific®, Austin, TX, USA)
equipped with a 100% Cyanopropyl silicone
capillary column (SPTM - 2340 60 m, 0.32 mm
ID, film thickness 0.25 mm). The oven
temperature was held at 140º C for 5 min and
subsequently increased at 4º C/min to 220º C.
Injector temperature was 250º C. Identification
of fatty acid methyl esters was based on
retention time of standard esters (Supelco® 37
Components FAME Mixture, Bellefonte, PA)
eluting from the capillary column. Peak areas
were integrated using chromatography data
software, and concentrations of each ester
were calculated as a percentage of the total
area of the chromatogram.
Atherogenic index (AI) and thrombogenic
index (TI) were calculated according to
Ulbricht and Southgate (1991).
3.7 Measurement of lipid oxidation
To evaluate oxidative stability, the cooked
samples were frozen at -18° C and analyzed
at 0, 15, 30, 60 and 90 days. Samples with

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0.01% butylated hydroxyanisole (BHA) were
used as positive reference in this assay.
Lipid oxidation of cooked beef patties was
monitored by measuring conjugated dienes
(CD) formation, peroxide value (PV) and
thiobarbituric acid reactive substances
(TBARS). Conjugated dienes were determined
according to Aubourg (1998). Pre-weighed
lipid samples were diluted with hexane and the
absorbance was measured at 233 nm in a
Beckman DU® 640B spectrophotometer
(Fullerton, CA). The hydroperoxides
conjugated dienes concentration was
expressed in milliliter per milligram of lipids.
The results were calculated as:
Where Bis the absorbance reading, V denotes
the volume (ml) of the solvent and w is the
mass (mg) of sample measured.
Determination of PV was conducted by the
IDF method (Shanta and Decker, 1994). 0.01
g of samples was dissolved in 9.9 ml
chloroform: methanol (70:30) solution and 0.05
ml of 30 % (w/v) ammonium thiocyanate were
added and mixed. Precisely 5 min after the
addition of 0.05 ml of ferrous chloride in 3.5 %
v/v hydrochloride acid to the reaction mixture,
the absorbance of the resulting red color was
measured at 501 nm against solvent solution
as blank. Results were expressed in meq O2
/kg of lipids. A modified thiobarbituric acid
reactive substances (TBARS) method was
used to evaluate the extent of lipid oxidation
(Ahn and Kim, 1998). One hundred milligrams
of lipids were taken, and the following
reactives were sequentially added: 100 µl
(BHA 36 g/l) and 2 ml of TBA/trichloroacetic
acid (TCA) solution (20 mM TBA in 150 g/l
TCA). The mixture was heated in a 90º C
water bath for 15 min and cooled at room
temperature. Afterwards, two milliliters of
chloroform were added and the mixture was
centrifuged at 1,000 g for 15 min. The
absorbance of the supernatant was measured
at 532 nm in an UV-Vis Beckman DU 640B®
spectrophotometer against a blank containing
0.1 ml H2O and 2 ml TBA/TCA solution.
Malonaldehyde (MAD) standard curves were
prepared by 1,1,3,3-tetramethoxypropane and
TBARS were expressed as mg/kg of MAD
equivalents of lipids.
3.8 Microbiological analyses
Ten grams of each sample were homogenized
with 90 ml of 0.1% sterile peptone water, and
appropriate serial dilutions were plated in
duplicate on plate count agar (PCA) at 30° C
for 48 h for total bacterial count. Results were
expressed as log number of colony forming
units per g of sample (log cfu/g).
3.9 Texture profile analysis
The cooked patties texture profile analysis
(TPA) was subjected to a two-cycle
compression test, as described by Pons and
Fiszman (1996), using a texture analyzer
(Stable Micro System TA-XT2i ®, Surrey,
England), equipped with a load cell of 4.5 kg.
Hardness, cohesiveness, gumminess,
instantaneous springiness, delayed
springiness and chewiness were obtained
using the available computer software. Three
different patties from each sample were
analyzed.
4. Statistical Analyses
The results were expressed as mean ±
standard deviation of the mean for the
formulations under study. All statistical tests
were performed using Statgraphics Plus for
Windows software package. Data collected
from proximate composition was analyzed by
using 5x2 factorial categoricaldesign, being
the analyzed factors: formulations (C; EP; EP
0.5% DSS; EP 1% DSS and EP2% DSS) and
heat treatment (raw and cooked). Differences
were considered significant at p<0.05 and the
Fisher’s Multiple Range Test were used for
comparison of mean values. One-way analysis
of variance was applied for the evaluation of
cooking properties, fatty acid profile, TPA and
microbiological analysis, considering each
formulation as a level. Data obtained from
oxidative stability parameters for each storage
time (0, 15, 30, 60 and 90 days) were
analyzed with one-way analysis of variance
(ANOVA) using the formulations as factor.
The experiment was replicated twice.
5. Results and Discussion
5.1 Proximate composition and fatty acid
profile of soybean sprouts
Proximate composition and alimentary fiber of
DSS used as additive for the preparation of
experimental burger are shown in Table 1.
Fatty acid composition is presented in Table 2.
(Values represent the mean value for n=3
replicates of DSS).
Table 1: Proximate composition of dehydrated
soybeans sprouts
Component g/100 g Mean± SD
Moisture 11.43±0.08
Protein 48.23±1.05
Fat 0.96±0.03
Carbohydrates 38.32±1.68

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Alimentary fiber 9.88±0.06
Ash 5.97± 4.85E-05
Sodium 0.32±0.20
All values are mean ± standard deviation of three
replicates
Table 2: Mean of fatty acid composition (% of fatty
acids)
Fatty acid % Total Mean± SD
C14:0 0.46±0.004
C16:0 21.50±0.143
C16:1 1.55±0.239
C17:0 0.55±0.008
C18:0 5.23±0.036
C18:1n9 9.65±0.050
C18:2n6 37.60±0.149
C18:3n3 12.47±0.004
C20:0 1.48±0.003
C22:0 5.34±0.006
C22:2n6 4.14±0.156
All values are mean ± standard deviation of three
replicates
5.2 Effect of soybean sprouts on proximate
composition of beef patties with partial
replacement of pork backfat by soybean oil
The mean percent moisture, protein, fat, ash,
carbohydrate contents and pH of raw and
cooked beef patties are given in Tables 3 and
4.
Moisture analysis of all patties showed
significant differences (p<0.05) between raw
and cooked samples, in addition the effect of
DSS on formulation was found to be
significant, improving moisture content in
cooking samples. Soybean sprouts addition
decreased moisture content in raw patties.
This is in agreement with others researchers
(Serdaroǧlu, 2006; Alakali et al., 2010; López-
López et al., 2010) who reported a decrease in
moisture content of beef patties formulated
with oat flour, bambara groundnut and
Wakame, probably because of increased solid
contents. However, adding soybean sprouts
resulted in a significant increase of moisture
content in cooked patties, this is because
addition of this ingredient reduced the drip and
evaporation (Alakali et al., 2010).
Dry soybean sprouts have a high content of
protein and carbohydrates (Table 1), which
help to improve rheological properties and
stability of emulsion, increasing retention of
moisture and fat.
The difference between raw and cooked
patties in terms of protein content was found to
be significant (p<0.05). The main effect for
protein content was presented by heat
treatment indicating that changes induced by
cookingare greater than formulation effect.
The protein content of cooked patties was
higher than the raw patties due to drip and
leaching decreased moisture content and
therefore increased protein content in cooked
patties. Alakali et al. (2010) reported the same
effect after cooking beef patties formulated
with bambara groundnut seed flours, and
López - López et al. (2011) informed that the
increase in protein levels after cooking is
consistent with the cooking loss.
Table 3: Mean values of proximate composition of patties (raw and cooked) formulated with different
concentrations of dry soybean sprouts
Samples Moisture Protein Carbohydrates Fat
Raw patties
C 63.03±0.38
Aa
22.65±0.56
Ab
0.59±0.02
Ab
19.17±0.05
Ac
EP 0% DSS 61.04±0.22
Aa
20.39±0.52
Ab
0.44±0.01
Aa
17.81±0.31
Aa
EP 0.5% DSS 59.12±0.62
Aa
19.27±0.07
Aa
0.66±0.01
Ac
18.32±0.11
Aa
EP 1% DSS 58.84±0.70
Aa
19.15±0.86
Aa
0.77±0.03
Ad
18.82±0.26
Aa
EP 2% DSS 59.53±0.22
Aa
19.65±0.02
Aab
0.86±0.08
Ae
17.13±0.05
Aa
Cooked patties
C 57.29±0.01
Ba
29.80±0.56
Bb
0.53±0.00
Ab
14.06±0.35
Bb
EP 0% DSS 58.94±0.61
Bb
32.09±2.49
Bb
0.54±0.01
Aa
9.66±0.07
Ba
EP 0.5% DSS 59.44±0.21
Bb
27.78±0.04
Ba
0.66±0.00
Ac
10.06±4.06E-04
Ba
EP 1% DSS 58.22±0.04
Bab
28.71±0.04
Ba
0.74±2.00E-04
Ad
10.37±0.02
Ba
EP 2% DSS 58.27±0.10
Bab
29.91±0.51
Bab
0.89±0.01
Ad
15.09±0.01
Bb
Results are presented as means ± standard deviation. Different letters (
A,a
) in the same column denote significant
differences among heat treatment and formulations respectively (p<0.05).
The higher addition of DSS, the higher the
carbohydrate content of both raw and cooked
patties, this could be due to the high content of
carbohydrates in soybean sprouts (Table 1).
For this parameter both factors exerted
significant effects (p<0.05).
As expected, fat content of cooked beef
patties decreased significantly after cooking,
even more when the fat was replaced by
soybean oil; both factors, heat treatment and
formulation exerted significant effect on this
parameter (p<0.05).Besides, fat content of
cooked patties tended to increase consistent

6. Ph ton 232
with the level of soybean sprouts added, due
to its high carbohydrates and proteins
contents, which improved the fat-absorption
capacity (Ali et al., 2011).
Ashes analysis of all samples showed
significant differences between heat treatment
and formulation (p<0.05) (Table 4).The higher
addition of DSS, the higher the ashes content
of cooked samples.
With respect to sodium content (Table 4),
although the values were between 800 and
1800 mg/100 g product, which is in
accordance with the common meat product
formulation (2% NaCl), this level increased
when the soybean sprouts were added in
increasing concentration to both raw and
cooked patties. This behavior could be
attributed to the sodium content of this
additive. Both factors, heat treatment and
formulation exerted significant effects
(p<0.05).
The pH values of uncooked and cooked
patties were significantly different (p<0.05) and
the cooking procedure slightly increased pH in
all treatments. This is in accordance with
Serdaroǧlu (2006) and López-López et al.
(2010).
Table 4: Mean values of ashes, sodium and pH content of patties (raw and cooked) formulated with different
concentrations of dry soybean sprouts
Samples Ashes Sodium pH
Raw patties
C 3.70±0.31
Ab
886.92±11.10
Aa
6.25±3.02E-03
Ab
EP 0% DSS 3.09±0.04
Aa
937.14±6.87
Ab
6.13±5.63E-03
Aa
EP 0.5% DSS 3.09±0.15
Aa
1077.01±7.08
Ac
6.39±2.25E-04
Ad
EP 1% DSS 2.95±0.04
Aa
1014.24±14.49
Ad
6.33±6.25E-04
Ac
EP 2% DSS 2.77±0.27
Aa
1418.69±15.09
Ae
6.37±2.02E-03
Ad
Cooked patties
C 2.87±0.02
Ba
1322.55±18.16
Ba
6.34±2.25E-04
Bb
EP 0% DSS 3.46±0.01
Bab
1424.26±9.54
Bb
6.22±8.00E-04
Ba
EP 0.5% DSS 3.50±0.25
Bb
1522.38±19.57
Bc
6.66±3.60E-03
Bd
EP 1% DSS 3.57±0.43
Bb
1470.67±9.55
Bd
6.49±1.22E-03
Bc
EP 2% DSS 3.68±0.17
Bb
1772.75±17.29
Be
6.65±4.22E-03
Bd
Results are presented as means ± standard deviation. Different letters (
A,a
) in the same column denote significant
differences among heat treatment and formulations respectively (P<0.05).
5.3 Effect of soybean sprouts on cooking
characteristics of beef patties with partial
replacement of pork backfat by soybean oil
Cooking characteristics of patties are given in
Table 5. Cooking yield, (p<0.05) were strongly
affected when the pork fat was replaced by
soybeans oil, decreasing the total weight of
patties.
On the other hand, the DSS incorporation
exerted a significant increase (p<0.05) in all
cooking parameters, being this effect higher
when the concentrations of soybean buds
raised up to 2%. This is in accordance with
Naveena et al. (2006), who observed that
addition of Ragi millet flour to chicken patties
resulted in better cooking yield, and with
Sanchez-Zapata et al. (2010), who indicated
that addition of tiger nut fiber causes desirable
changes in the cooking characteristics of the
pork burgers. This behavior could be attributed
to the high ability of the DSS to keep moisture
and fat in the matrix.
Dimensional changes are important in
maintaining quality standards of burgers linked
to potential negative reactions of consumers
due to the negative image of excessive water
Table 5: Mean cooking properties and dimensional changes of beef patties by replacing pork back fat with soy oil
with different concentrations of DSS
Sample Cooking
yield (%)
Fat retention
(%)
Moisture
retention (%)
Diameter
reduction
(%)
Thickness
contraction (%)
Shrinkage
(%)
C 78.23±0.15
b
56.71±4.46
c
73.00±4.28
ab
19.12±5.09
a
12.22±4.71
a
20.24±0.26
a
EP 0% DSS 74.64±0.00
a
40.33±0.07
a
72.08±2.19
a
23.17±0.79
a
14.17±4.32
a
21.67±1,76
a
EP 0.5% DSS 76.66±0.36
b
41.64±0.01
ab
77.85±2.04
bc
23.39±0.44
a
16.39±0.69
a
21.76±0.06
a
EP 1% DSS 76.92±0.28
b
44.11±0.09
b
78.86±0.74
c
22.01±1.92
a
20.38±0.14
a
20.38±0.54
a
EP 2% DSS 80.46±0.46
c
71.73±0.36
d
79.07±0.74
c
22.91±0.39
a
20.83±1.92
a
22.36±1.31
a
Results are presented as means ± standard deviation. Different letters in the same column denote significant
differences among formulations (P<0.05).

7. Ph ton 233
addition. Therefore, the impacts of added
ingredients in the dimensional changes were
evaluated. The results obtained in this case,
showed that although pork fat was replaced
and DSS were added to beef patties,
significant changes in dimensional
characteristics (diameter reduction, thickness
and shrinkage were not observed
(p>0.05).This is probably due to the binding
and stabilizing properties of soybean buds,
which maintain the meat particles together
avoiding changes in the shape of the products
(Choi et al., 2012).
5.4 Effect of soybean sprouts on fatty acids
profile of cooked beef patties with partial
replacement of pork fat by soybean oil
Vegetable oils with a high content in PUFA
such as corn, sunflower, cotton seed, and
soybean oil, have been used to substitute the
fat content in cooked meat burgers to increase
the PUFA/SFA ratio as a consequence of a
higher content in linoleic acid and α-linolenic
acid, which exerts a relevant influence on the
nutritional quality of the products (Rodríguez-
Carpena et al., 2012).
In this study, the replacement of pork fat by
soybean oil in raw patties formulation
increased n3 PUFA of short chain content
from 1.65% to 4.55%, and Σ PUFA from 23%
to 45% (Table 6 and Table 7). Moreover, as a
consequence of replacement, SFA acids were
reduced too; therefore the PUFA/SFA
relationship increased around three times, and
the n6/n3 ratio was maintained below 10.
The improvement in raw products fatty acid
composition was affected by the cooking
process, decreasing 21% and 18% the linoleic
acid content and the alpha-linolenic content,
respectively. This loss could be explained by
the lower melting point of polyunsaturated fatty
acids and the poor retention capacity of the
meat matrix used. However, in cooked patties
when DSS was added the loss of these fatty
acids were minimized, further improving the
PUFA/SFA relationship, and maintaining n6/n3
ratio below 10 (Figure 1 A and B).
Products obtained in this ways, were healthier
due to the higher contents of unsaturated
acids and essential fatty acids. This is in
accordance with López-López et al. (2011),
who reported a significant increase of the
PUFA/SFA ratio in some formulations of
patties with olive oil and Wakame seaweed
added. Salcedo-Sandoval et al. (2013)
informed that the n6/n3 ratio remained near 10
in meat products with reduced fat content
(pork back fat replaced by emulsified oil-in-
konjac matrix).
Table 6: Fatty acid profile of raw patties formulated with different concentrations of dry soybean sprouts (DSS) in
g/100g dry product
Fatty acid Raw Samples
C EP0% DSS EP 0.5 % DSS EP 1% DSS EP 2% DSS
(14:0) 1.23±0.01
c
0.62±0.00
a
0.61±0.01
a
0.73±0.03
b
0.72±0.01
b
(16:0) 21.49±0.15
d
15.30±0.14
a
15.09±0.16
a
16.12±0.03
c
15.62±0.03
b
(16:1) 1.90±0.21
b
0.96±0.01
a
0.93±0.02
a
1.11±0.11
a
0.99±0.01
a
(17:0) 0.53±0.17
b
0.30±0.03
a
0.29±0.01
a
0.34±0.01
ab
0.34±0.00
ab
(17:1) 0.48±0.21
d
0.23±0.03
b
- 0.28±0.01
c
0.23±0.03
b
(18:0) 11.77±0.37
b
7.87±0.08
a
7.88±0.06
a
8.29±0.21
a
8.29±0.17
a
(18:1)t n9 0.41±0.11
c
- - 0.20±0.06
b
0.26±0.11
bc
(18:1)c n9 39.23±0.20
d
29.72±0.08
b
29.08±0.07
a
31.13±0.03
c
29.53±0.08
b
(18:2)c n6 12.42±0.35
a
37.09±0.07
d
36.77±0.01
d
33.24±0.21
b
35.93±0.04
c
(20:0) - 0.26±0.02b
c
0.26±0.01
bd
0.22±0.02
b
0.30±0.01
c
α (18:3) n3 1.65±0.35
a
4.55±0.20
c
4.55±0.28
b
4.10±0.04
b
4.47±0.10
b
(18:2) CLA 8.90±0.45
e
3.11±0.01
c
4.60±0.07
b
4.23±0.07
b
3.32±0.03
a
ΣSFA 35.01±0.71
d
24.35±0.28
ab
24.12±0.25
a
25.70±0.30
c
25.26±0.23
bc
ΣMUFA 42.02±0.74
d
30.91±0.17
ab
30.01±0.01
a
32.73±0.04
c
31.01±0.05
b
ΣPUFA 22.96±1.16
a
44.75±0.37
cd
45.87±0.30
d
41.57±0.33
b
43.72±0.17
c
Results are presented as means ± standard deviation. Means in the same row with different letters are
significantly differentamong formulations (P<0.05). N.D.: not detected.
The AI assesses the risk of atherosclerosis
and the TI is an indication of the potential
aggregation of blood platelets, so very low
values of these indexes are recommended for
a healthy diet.
Regarding AI and TI indices, the values
calculated using Ulbricht and Southgate
(1991) equation from samples partially
replaced by soybean oil were lower than the
controls (p<0.001) for both studied factors
This is according to Salcedo-Sandoval et al.
(2013), who reported that the samples

8. Ph ton 234
containing only animal fat presented greater
values than frankfurters with healthier oil
replacement. Furthermore, DSS addition did
not affect these indices in both raw and
cooked patties. This behavior was similar to
the results found by López-López et al. (2011)
when seaweeds were added to beef patties
whose pork fat was partially or totally replaced
by olive oil in water emulsion.
Figure 1: Fatty acid profile of cooked meat products, (A) without soybean oil and (B) with soybean oil
SI: C 19:0, Internal Standard; RT: 17.373 C14:0; RT: 20.965 C16:0; RT: 22.087 C16:1; RT: 22.670 C17:0; RT:
23.744 C17:1; RT: 24.483 C18:0; RT: 25.192 C18:1 n9 t; RT: 25.498 C18:1 n9 c; RT: 26.947 C18:2 n6; RT:
28.673 C18:3 n3; RT: 30.112 CLA C18:2 n6
Table 7: Fatty acid profile of cooked patties formulated with different concentrations of dry soybean sprouts
(DSS) in g/100g dry product
Fatty acid
Cooked Samples
C EP EP DSS 0.5% EP DSS 1% EP DSS 2%
(14:0) 1.30±0.51
b
0.78±0.08
ab
0.75±0.01
ab
0.71±0.06
ab
0.65±0.06
a
(16:0) 21.69±1.75
b
16.70±0.21
a
16.48±0.13
a
16.52±0.35
a
15.75±0.14
a
(16:1) 1.97±0.42
b
1.26±0.01
a
1.12±0.00
a
1.12±0.11
a
0.95±0.04
a
(17:0) 0.54±0.11
b
0.36±0.01
a
0.33±0,07
a
0.33±0.04
a
0.31±0.03
a
(17:1) 0.50±0.16
b
0.33±0.06
ab
0.26±0.01
a
0.25±0.01
a
0.17±0.08
a
(18:0) 11.77±1.53
b
8.41±0.07
a
8.68±0.13
a
8.46±0.04
a
8.21±0.10
a
(18:1)t n9 0.38±0.16
a
0.23±0.01
a
0.18±0.14
a
0.22±0.13
a
0.33±0.10
a
(18:1)c n9 39.96±1.27
d
32.90±0.35
c
31.08±0,13
ab
31.26±0.38
b
29.49±0.20
a
(18:2)c n6 12.00±2.87
a
29.16±0.35
b
33.39±0.20
c
33.46±0.14
c
33.96±0.04
c
(20:0) NDa 0.18±0.01
b
0.22±0.02
bc
0.20±0.01b
c
0.27±0.03
c
α (18:3) n3 1.74±0.33
a
3.70±0.30
b
4.14±0.18
b
4.15±0.06
b
4.07±0.08
b
(18:2) CLA 8.21±1.03
c
5.98±0.03
b
3.36±0.04
a
3.32±0.11
a
5.84±0.16
b
Σ SFA 35.25±0.19
c
26.42±0.17
b
26.46±0.14
b
26.22±0.20
b
25.19±0.33
a
ΣMUFA 42.80±2.01
c
34.72±0.61
b
32.64±0.28
ab
32.85±0.64
ab
30.94±0.42
a
ΣPUFA 21.95±4.93
a
38.84±1.34
b
40.86±0.32
b
40.93±0.31
b
43.89±0.27
b
n3 1.74±0.33
a
3.70±0.30
b
4.11±0.08
b
4.15±0.06
b
4.07±0.08
b
n6 20.21±3.9
a
35.14±1.05
b
36.75±0.24
bc
36.78±0.25
bc
39.82±0.19
c
n9 39.96±1.27
d
32.91±0.35
c
31.08±0.13
ab
31.26±0.38
b
29.49±0.20
a
PUFA/SFA 0.62±0.05
a
1.47±0.03
b
1.54±0.01
bc
1.56±0.02
c
1.74±0.03
d
n6/n3 11.61±0.07
c
9.52±0.48
ab
8.94±0.13
a
8.86±0.06
a
9.78±0.16
b
AI 0.41±0.02
c
0.27±0.00
b
0.27±0.00
ab
0.26±0.01
ab
0.25±0.00
a
TI 0.94±0.00
d
0.56±0.01
c
0.55±0.01
bc
0.54±0.00
b
0.52±0.00
a
Results are presented as means ± standard deviation. Means in the same row with different letters are
significantly differentamong formulations (p<0.05). N.D.: not detected.
5.5 Effect of soybean sprouts on oxidative
stability of beef patties with partial replacement
of pork fat by soybean oil
Modern life led to an increase of production
and consumption of precooked frozen meat
products. However, processed cooked meat
products are more susceptible to lipid
peroxidation during chilled and frozen storage,

9. Ph ton 235
due to the fact that the heating process leads
to a dramatic increase of oxidative reactions of
lipids in meat, which cause a warmed-over-
flavour (WOF) (Bastida et al., 2009).
The formation of Conjugated Dienes (CD)
occurs parallel to the production of
hydroperoxides (measured as PV) and take
place at the early stages of lipid oxidation
(Frankel, 1996); later, these compounds
decompose into secondary products such as
aldehydes and ketones. The increase of PV in
cooked samples may result from catalysis of
intracellular compounds due to the destruction
of self-structures by NaCl and processing
(Roginsky and Barsukova, 2001). TBARS
values represent the content of secondary lipid
oxidation products mainly aldehydes (or
carbonyls), which contribute to off-flavors in
oxidized meat and meat products.
Moreover, the oxidative stability decreases
more when the lipid composition of these
products was improved with polyunsaturated
fatty acids; thus, antioxidants are usually used
to prevent lipid oxidation. Earlier, DSS had
shown to possess a moderate antioxidant
effect in other matrixes, therefore to verify if
DSS added to enriched meat patties exert this
effect, conjugated dienes, hydroperoxides and
aldehydes formed during frozen storage were
continuously monitored. These parameters
(CD, PV and TBARS) were determined on
cooked meat patties and are presented in
Table 8. Using the CD determination as lipid
oxidation indicator, DSS concentrations did not
show antioxidant activity until day 30, although
the CD formation at 60 and 90 days was lower
than control for 0.5% and 1% of this additive
(p<0.05), while the 2% concentration exerted
pro-oxidant effect at 30 days.
Regarding hydroperoxide and TBARS
formation, 0.5 % and 1% concentration of DSS
added exerted antioxidant effect at 90 days
(p<0.05); however, 2% concentration was only
effective to inhibit lipid peroxidation at 15 days
of frozen storage of cooked patties. This is in
accordance with Peña-Ramos and Xiong
(2003) and Sánchez-Alonso et al. (2007), who
reported the same effect with soy protein
isolates and hydrolyzates in cooked pork
patties during refrigerated storage at 7 days of
storage, and with grape dietary fiber at 30
days of frozen storage in minced fish
respectively. In summary, 0.5% and 1% DSS
concentrations exerted antioxidant effect at 90
days with reduction percentages of lipid
oxidation with respect to control without
antioxidant of 64% and 51% for PV, and 58%
and 54% for TBARS respectively, while the
2% DSS concentration showed prooxidant
effect at 30 days of frozen storage.
Considering the fat retention capacity and the
oxidative stability, the patties added with 1%
dehydrated soybean sprouts were selected for
the following analysis.
Table 8: Average values of CD, PV and TBARS in cooked patties with different DSS concentration and BHA
treatment during 90 days at frozen storage
Samples C
EP
0.5% DSS
EP
1% DSS
EP
2% DSS
EP
0.01% BHA
Conjugated Dienes (ml CD/ mgsample)
Day 0 0.52±0.00
b
0.48±0.02
b
0.52±0.01
b
0.54±0.01
c
0.42±0.01
a
Day 15 0.47±0.01
b
0.49±0.01
b
0.46±0.00
b
0.47±0.01
b
0.35±0.00
a
Day 30 0.52±0.02
b
0.52±0.02
b
0.52±0.02
ab
0.75±0.01
c
0.46±0.01
a
Day 60 0.61±0.01
c
0.52±0.01
b
0.55±0.01
bc
0.82±0.03
d
0.48±0.01
a
Day 90 0.62±0.01
c
0.52±0.02
b
0.60±0.05
bc
0.89±0.03
d
0.49±0.01
a
Peroxide Value (meq O2/kg sample)
Day 0 23.87±3.38
e
4.52±0.04
b
10.80±0.06
c
12.78±0.00
d
0.88±0.13
a
Day 15 35.78±1.17
e
20.85±0.07
d
13.42±0.01
b
15.58±0.00
c
2.08±0.02
a
Day 30 34.16±0.00
c
23.33±0.72
b
22.91±0.06
b
61.66±5.13
d
3.11±0.01
a
Day 60 46.16±1.51
c
22.76±0.25
b
25.38±0.06
b
69.57±4.88
d
3.96±0.10
a
Day 90 58.17±4.27
d
20.989±2.50
b
28.55±0.36
c
77.47±3.16
e
4.80±0.00
a
TBARs (mg MAD/ kg dry matter)
Day 0 2.55±0.00
c
1.64±0.00
b
2.23±0.01
bc
4.88±0.18
d
0.60±0.01
a
Day 15 3.45±0.10
c
3.38±0.02
c
2.98±0.01
bc
2.73±0.02
b
1.57±0.10
a
Day 30 3.57±0.33
b
3.34±0.16
b
3.07±0.10
b
5.99±0.23
c
0.80±0.33
a
Day 60 5.95±0.16
c
3.55±0.07
b
3.32±0.15
b
8.18±0.41
d
0.79±0.17
a
Day 90 8.33±0.65
c
3.47±0.03
b
3.80±0.01
b
10.36±0.01
d
0.80±0.01
a
Results are presented as means ± standard deviation. Different letters in the same row denote significant
differencesamong formulations (p<0.05).

10. Ph ton 236
5.6 Texture profile analysis (TPA) of beef
patties with partial replacement of pork fat by
soybean oil
Table 9 shows the effects of replacing fat
content with soybean oil and soybean sprouts
as natural antioxidants on the textural
properties of selected cooked beef patties.
Significant changes in hardness,
cohesiveness, chewiness and delayed
elasticity were recorded. The hardness of
cooked meat patties significantly decreased
(p<0.05), this is in accordance with López-
López et al. (2010), who reported that
Wakame addition to low-fat beef patties
softened the products.
Table 9: Effects of substituting pork back fat with soybean oil and 1% DSS as natural antioxidant on texture
profile analysis of meat products
Samples Hardness (N) Cohesiveness Instantaneous
elasticity
Delayed
elasticity
Chewiness
(N)
Gumminess
Control 15.02±2.40
b
0.59±0.01
a
0.47±0.04
a
0.94±0.23
b
6.03±1.55
a
7.07±1.12
a
EP1% DSS 10.83±0.40
a
0.67±0.03
b
0.56±0.02
b
0.76±0.12
ab
6.17±1.48
a
6.93±0.33
a
EP 12.64±2.51
ab
0.61±0.04
a
0.55±0.01
b
0.69±0.02
a
5.29±1.23
a
7.69±1.77
a
Results are presented as means ± standard deviation. Different letters in the same column denote significant
differences among formulations (P<0.05).
This could be due to the fact that the
tridimensional network of the seaweed
insoluble dietary fiber was less distributed in
the meat matrix, showing less opportunity to
interact with it. The chewiness and gumminess
of enriched and DSS added patties were
similar when compared to the control (p>0.05),
while the cohesiveness and instantaneous
elasticity were higher than the samples without
antioxidants (p<0.05). The variations in texture
among the enriched samples and control may
have also been due to the differences in the
physicochemical characteristics of the lipid
phase (solid pork fat versus soybean oil).
5.7 Microbiological analysis of beef patties
with partial replacement of pork fat by soybean
oil
The patties elaboration process involves a lot
of manipulation and therefore could affect the
hygiene in these products; hygiene being
important due to its connection with enteric
diseases (Karr et al., 1996). The total viable
counts for raw and cooked samples did not
exceed 4 to 4.3 log cfu/g. Similar counts have
been reported by López-López et al. (2010);
Naveena, et al. (2006) and Das et al. (2008).
Besides, no changes were observed in
microbial populations of samples during the
entire assay, then it was concluded that in the
given experimental conditions, all products
were microbiologically stable and safe to eat.
Conclusions
Protein and carbohydrate content, moisture
and fat retention were improved when dry
soybean sprouts were added to soybean oil
enriched beef patties. The cooking loss was
minimized and the shape and size of the
samples were not affected. The 0.5% and 1%
DSS concentrations showed antioxidant effect
at 90 days of frozen storage, while the 1% was
the most effective to retain healthier lipids. The
selected samples with 1% concentrations
presented a softer texture and an acceptable
microbiological quality.
Replacement of pork fat with soybean oil
added with 1% soybean sprouts as natural
additive resulted in more nutritional and
healthier meat products. In addition,
development of functional foods with soybean
sprouts as food ingredient opens up new
possibilities to their use as additive.
Research Highlights
Soybean oil improved fatty acid profile of
beef patties.
Dry soybean sprouts can be used as natural
antioxidant to protect beef patties against
lipid oxidation.
Quality characteristics of beef enriched
patties were evaluated.
1% of DSS was most effective to retain and
maintain healthier lipids
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