LWT - Food Science and Technology 46 (2012) 548e555

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LWT - Food Science and Technology

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Total phenolic and total flavonoid content, antioxidant activity and sensory
evaluation of pseudocereal breads
Joanna Chlopicka a, Pawel Pasko a, *, Shela Gorinstein b, Aneta Jedryas a, Pawel Zagrodzki a, c
a
Department of Food Chemistry and Nutrition, Medical College, Jagiellonian University, 9 Medyczna Street, 30-688 Krakow, Poland
b
Institute for Drug Research, School of Pharmacy, The Hebrew University e Hadassah Medical School, Jerusalem 91120, Israel
c
Henryk Niewodniczan  ski Institute of Nuclear Physic, Department of Nuclear Physical Chemistry, 152 Radzikowskiego Street, 31-342 Krakow, Poland

a r t i c l e i n f o a b s t r a c t

Article history: The aim of the study was to investigate the effect of adding (in two different doses 15% and 30%)
Received 8 April 2011 pseudocereal (buckwheat, amaranth and quinoa) flour on the antioxidant properties and sensory value
Received in revised form of breads. Buckwheat flour had the highest phenolic content (7.25
0.23 mg/g dw). The content of total
10 November 2011
flavonoids in flours was about 2e4 fold higher when compared to breads. The addition of buckwheat
Accepted 13 November 2011
flour to wheat bread, particularly in higher dose, was more effective in enhancing antioxidant activity, as
evaluated by means of FRAP and DPPH, which increased by 2.36 fold, and 3.64 fold respectively, in
Keywords:
comparison with other pseudocereal flours (amaranth, quinoa), which caused, in higher doses, the
Pseudocereal
Bread
changes of above parameters within the ranges 1.20e1.79 fold, and 0.60e1.71 fold. Analysis of sensory
Antioxidant activity results of breads showed that addition of buckwheat flour to the dough might improve subjective
Sensory quality properties of bread and increase acceptable quality attributes such as taste, colour or odour. All these
observations suggest that addition of buckwheat flour into bread can improve antioxidant as well as
sensory properties of bread. Bread fortified with pseudocereal flours, and especially with buckwheat
flour, may be placed on the market as a functional food.
Ó 2011 Elsevier Ltd. All rights reserved.

1. Introduction of traditional type of food, i.e. “slow food”, which is now becoming
more popular. Increasing consumption of this kind of bread in our
Whole pseudocereal grains such as buckwheat, amaranth and daily menu can improve antioxidant potential of our diet because it
quinoa are rich in a wide range of compounds e.g. flavonoids, is known that the pseudocereals’ grains are rich in different anti-
phenolic acids, trace elements, fatty acids and vitamins with known oxidant compounds (Lin, Liu, Yu, Lin, & Mau, 2009; Pasko, Sajewicz,
effects on human health (Gorinstein et al., 2008; Kalinova & Gorinstein, & Zachwieja, 2008). Pseudocereals can provide benefi-
Dadakova, 2009; Li & Zhang, 2001; Tomotake et al., 2007). Whole 
cial health effects (Christa & Sorel-Smietana, 2008; Martirosyan,
grain products consumption have been associated with reduced Miroshnichenko, Kulakova, Pogojeva, & Zoloedov, 2007), therefore
incidence of diseases such as cancer (Chan, Wang, & Holly, 2007; bread with addition of buckwheat, amaranth or quinoa flour, as
Slavin, 2004) cardiovascular disease (Jacobs & Gallaher, 2004; a staple product could diversify ordinary (daily) model of nutrition.
Mellen, Walsh, & Herrington, 2008), high blood pressure (Behall, That is why, the sensory value of bread is very important, because
Scholfield, & Hallfrisch, 2006; Flint et al., 2009) and diabetes taste, smell, and flavour of bread significantly influence consumer
(Lutsey et al., 2007; Qi & Hu, 2007; Rave, Roggen, Dellweg, Heise, & preferences of cereal products.
tom Dieck, 2007). Therefore, increased consumption of grain The sensory characteristics, chemical, rheological and storage
products has been recommended, and cereals products should be properties of breads from various species of wheat flour or breads
the main part of the daily menu (Richardson, 2003). The pseudo- from different portions of other cereal flours, have been thoroughly
cereals are not often used in technology of bread but they can be investigated. It was observed, that the additions of some flours
useful in dietotherapy of celiac disease (Thompson, 2001). Home- having valuable nutrient profile to poor quality common soft wheat
made breads containing pseudocereals could be an important part improved the breadmaking properties, extended the shelf life of
the corresponding product, made crumb more softer, and lowered
firming rate. The nutritive properties of breads (e.g. the protein
* Corresponding author. quality and quantity, dietary fibre content, and unsaturated fats
E-mail address: paskopaw@poczta.fm (P. Pasko). profile) were also enhanced (Raffo et al., 2003). However, little is

0023-6438/$ e see front matter Ó 2011 Elsevier Ltd. All rights reserved.
doi:10.1016/j.lwt.2011.11.009
 J. Chlopicka et al. / LWT - Food Science and Technology 46 (2012) 548e555 549

known about antioxidant effect (Alvarez-Jubete, Wijngaard, Arendt, of eight stages, i.e.: 15 min mixing, 15 min growing without raising
& Gallagher, 2009) and sensory characteristics of pseudocereal temperature, 10 min mixing, 15 min growing without raising
breads, especially about breads with addition of amaranth and temperature, 45 min growing in low temperature (35  C), 10 min
quinoa flour (Alvarez-Jubete, Auty, Arendt, & Gallagher, 2009; mixing, 15 min growing in medium temperature (45  C), 50 min
Nikoli
c, Sakac, & Mastilovic, 2011; Schoenlechner, Drausinger, baking. In case of buckwheat breads, time of baking had to be
Ottenschlaeger, Jurackova, & Berghofer, 2010). Therefore, the aim prolonged because in shorter time slack-baked bread was obtained.
of our study was to investigate the effect of addition of pseudo- Detailed data about time of baking and weight of bread after baking
cereal flour on the antioxidant properties and sensory value of were also collected in Table 1.
breads.
2.3. Extracts preparation
2. Materials and methods
After baking, breads were allowed to cool down to room
2.1. Materials temperature for 3 h. Subsequently, the breads were sliced (slices
about 1.5 cm thick), and kept frozen (20  C) until analysis. After
The flour used in the formula of control bread was wheat flour thawing, the slices were dried and then manually crumbed,
(Królowa Ma˛ k Tortowych, PPHU M1ynpol Sp. j. Gromadka, Krzy- grounded in traditional stone mortar and sieved through at 2 mm
_
zowa, Poland), type 400. The rest of breads were baked with sieve to obtain bread powder. Powdered samples of breads (2 g)
addition of buckwheat flour (Futuro c.m., Kraków, Poland), were extracted for 2 h with 40 mL of solvent consisting of
amaranth flour (PHU Szar1at s.c., qomza, _ Poland) or quinoa flour methanol, 0.16 mol/L hydrochloric acid and water, mixed in
(Futuro c.m., Kraków, Poland). The pseudocereal flours were added proportion 8:1:1, respectively. The extracts were separated by
in two different doses 15 g/100 g and 30 g/100 g, respectively. All of decantation and the residues were extracted again with 40 mL of
used flours were purchased from a local shop with “slow food” in 70 g/100 g acetone for 2 h. The initial methanol extracts were
Krakow, Poland. The other ingredients used in the formula of dough added to prepare mixture, which was subsequently decanted,
were: sugar (Kryszta1, Krajowa Spó1ka Cywilna S.A., Siennica centrifuged and stored in darkness in a freezer in temperature
Nadolna, Poland), iodinated salt (Tesco/Poland/Sp. ZO.O., Presov, of 20  C.
Slovak Republic), yeast (Dr Oetker Poland Sp. ZO.O., Gdan  sk,
Poland) and distillated water. Distilled water was used to eliminate
possible influence of some metals, e.g. iron and copper, on the 2.4. Determination of total phenols
evaluation of antioxidant activity, as it is well known that active
compounds in produced breads such as flavonoids and other Total phenols (TP) were determined colorimetrically using
polyphenols are vulnerable to oxidation in the presence of tran- FolineCiocalteu reagent, as described previously (Pasko et al.,
sient metals and ambient oxygen. 2009). Total phenols assay was conducted by mixing 2.7 mL of
De-ionized water for chemical analyses 18 Mohm cm was ob- de-ionized water, 0.3 mL of extracts, 0.3 mL 7 g/100 g Na2CO3 and
tained from Milli Ro & Q water purification system (Millipore, 0.15 mL FolineCiocalteu reagent. Absorbance of mixture was
Warsaw, Poland); methanol, acetone, hydrochloric acid 36 g/100 g, measured at 725 nm using the spectrophotometer Jasco UV-530
ferric chloride (FeCl3), aluminium chloride hexahydrate, sodium (Medson, Paczkowo, Poland). A standard curve was prepared with
nitrite, sodium hydroxide, sodium acetate and acetic acid were gallic acid. Final results were given as gallic acid equivalents (GAE).
purchased from Chempur, Piekary Sla  ˛ skie, Poland. 1,1-diphenyl-2-
picrylhydrazyl (DPPH), 6-hydroxy-2,5,7,8-tetramethyl-2- 2.5. Determination of total flavonoids
carboxylic acid (Trolox), FolineCiocalteu reagent, catechin and
gallic acid were purchased from Sigma Poznan  , Poland. All reagents Total flavonoid content was determined by a colorimetric
were of analytical grade. method as described previously (Gorinstein et al., 2007). Briefly,
0.25 mL of the 80 g/100 g methanolic extract was diluted with
2.2. Bread making 1.25 mL of distilled water. Then 75 mL of 5 g/100 g NaNO2
solution was added to the mixture, and after 6 min 150 mL of
The materials for dough-making were mixed according to the 10 g/100 g AlCl3$6H2O solution was added. The mixture was
formula proportions listed in Table 1. In this experiment, standard allowed to stand for 5 min and next 0.5 mL of 1 mol/L NaOH was
baking procedures were not used because the aim of this work was added and the total was made up to 2.5 mL with distilled water.
to present results of home-made breads baked using popular home The solution was mixed well and the absorbance was measured
bread oven. All types of baked breads were tin breads. The breads immediately against the blank at 510 nm using a spectropho-
were baked in the oven Alaska BM 2600 (METRO Group, Düsseldorf, tometer Jasco UV-530. The results were expressed as mg of
Germany) in temperature 180  C. Cycle of bread making composed catechin equivalents.

Table 1
The formulations of dough.

Type of bread Wheat Pseudocereal Salt Sugar Yeast Water Time of procedure Weight of bread
flour [g] flour [g] [g] [g] [g] [mL] [h min] after baking [g]
Control bread 500 e 10 10 5 350 2.55 780
Amaranth bread 15 g/100 g 425 75 10 10 5 350 2.55 780
Amaranth bread 30 g/100 g 350 150 10 10 5 350 2.55 787
Buckwheat bread 15 g/100 g 425 75 10 10 5 350 3.25 734
Buckwheat bread 30 g/100 g 350 150 10 10 5 350 3.25 748
Quinoa bread 15 g/100 g 425 75 10 10 5 350 2.55 781
Quinoa bread 30 g/100 g 350 150 10 10 5 350 2.55 778
550 J. Chlopicka et al. / LWT - Food Science and Technology 46 (2012) 548e555

2.6. Determination of FRAP activity asked to assess the breads for overall quality, based on the colour,
odour and consistency. The students were also asked about taste of
FRAP (Ferric Reducing Ability of Plasma) assay was carried out bread. They could choose: interesting, tasty, natural, strange taste,
according to Benzie and Strain (1996), and modified to 48-well vapid, not to eat, bad taste, gummy, crusty, little pronounced,
plates and automatic reader (Synergy-2, BioTek/USA) with syringe sweet, salty, tart, difficult to determine, delicate, milky.
rapid dispensers. Briefly, the oxidant in the FRAP assay (reagent
mixture) consisted of ferric chloride solution (20 mmol/L), TPTZ 2.9. Statistical analysis
solution (10 mmol/L TPTZ in 40 mmol/L HCl) and acetate buffer
(pH ¼ 3.6) in a proportion of 5:5:10, respectively, and was freshly Results of biochemical analyses are given as means
SD based
prepared. To each plate, 0.4 mL of acetate buffer (pH 3.6) was on four measurements for each sample of flour and bread. Where
dispensed, followed by 50 mL of sample, standard or blank. The appropriate, the data were tested by one-way ANOVA, followed by
plate was conditioned at the temperature of 37  C for 2 min, and Tukey post hoc test. For sensory parameters the median values and
then 0.2 mL of reagent mixture was added and shaken for 30 s; confidence intervals of medians were calculated (Bland, 2000). The
afterwards, absorbance at 593 nm was measured with kinetic mode possible differences between various breads, in respect to sensory
for 15 min. The final results were expressed as mg Trolox/100 g DW parameters, were analysed using KruskaleWallis test. Dunn’s post-
(dry weight). test was used to reveal the differences between the paired brands of
breads. Differences with p < 0.05 were considered to be statistically
2.7. Determination of DPPH radical scavenging activity significant. Pearson correlation coefficients were calculated for
pairs of biochemical parameters. Statistical calculations were
DPPH radical scavenging activity was measured according to the carried out using the commercially available packages Statistica
method of Yen and Chen (1995) with modification (Pasko et al., v.5.1 (StatSoft, Ic., Tulsa, USA), and GraphPad Prism v. 3.02
2009). For measurement of sample scavenging activity 0.4 mL of (GraphPad Software, San Diego, USA).
methanolic acetate buffer was added to the cuvettes containing the
increasing volumes of sample (e.g. 0, 0.1, 0.2, 0.3, 0.45, 0.6 mL) with 3. Results and discussion
adequate volumes of methanol to make total volume of 1 mL.
Acetate buffer was made from 0.2 mol/L solutions of sodium acetate 3.1. Phenolic contents of flour and breads
and acetic acid in methanol mixed at the volume ratio 7.9:2.1. The
pH of the buffer was 5.2. 1 mL of DPPH stock solution (12 mg DPPH The phenolic contents of the four kinds of flour and seven breads
was dissolved in 100 mL of methanol; absorbance 1.3) was added to were expressed as mg gallic acid per gram of dry weight (Table 2).
each cuvette, then absorbance was measured after 24 h. The Buckwheat flour had the highest phenolic content (7.25
0.23 mg/
absorbance of the resultant solution was determined using Jasco g dw) and the next one was wheat (6.96
0.11 mg/g dw). Amaranth
UV-530 spectrometer (Japan) at 514 nm. The total antioxidant and quinoa flour had the lowest phenolic content (2.71
0.1 mg/
capacities (TAA) were estimated as Trolox equivalents (TEAA) by g dw and 2.8
0.1 mg/g dw, respectively) and the differences
interpolation to 50% inhibition (TEAA50). between them and the former two were statistically significant.
Our results for phenolic content in buckwheat flour were in
2.8. Sensory evaluation between those given by Sensoy, Rosen, Ho, and Karwe (2006). They
observed lower content of total phenolics in buckwheat white, raw
The sensory evaluation was carried out on the breads 12 h after flour, but higher content of total phenolics of buckwheat dark, raw
baking. The 31 consumers (20 women; 11 men) aged 19e24 years flour in comparison with phenolic content in our buckwheat flour.
old completed the questionnaire. They were recruited among Consistently with the above results, the content of phenols in
students of Faculty of Pharmacy in Krakow. The samples were sliced breads was highest in breads baked with 30 g/100 g addition of
into equal sizes (2 cm  2 cm) before serving to the respondents on buckwheat flour (2.65
0.10 mg/g dw). However, the second
coloured coded plates. The consumers made hedonic evaluation of highest result was, surprisingly, for the same dose of amaranth
the samples. The scorecard was developed with 10-point category flour (2.61
0.04 mg/g dw), followed by bread with 30 g/100 g
scale (disliked ¼ 0; extremely liked ¼ 10) (Wronkowska, addition of quinoa flour (2.54
0.11 mg/g dw) and with 15 g/100 g
Troszyn 
 ska, Sorel-Smietana, & Wo1ejszo, 2008), each testers was addition of buckwheat flour (2.1
0.08 mg/g dw). Remaining

Table 2
Mean values (
SD, n ¼ 4) of total phenolic content, total flavonoids content, FRAP and DPPH for flours and breads under study (results marked with the same letter in upper
index within each column differ significantly).

Material tested Total phenolic Total flavonoids TEAC FRAP [mg TEAC DPPH [mmol
content [mg/g dw] content [mg/g dw] Trolox/100 g dw] Trolox/kg dw]
Wheat flour 6.96
0.1a,b,p,l 70
11a,b,f 158.3
3.0a,b,c,z 3.95
0.11b,h,k,c
Buckwheat flour 7.25
0.2p,c,d,m,n 153
12a,c,d,g,h 214.9
3.5a,e,f,u,w 8.80
0.52c,h,i,d,3
Amaranth flour 2.71
0.1a,c,o 65
8c,e,i,j 38. 6
1.2b,e,h,x,y 3.60
0.34i,m,4
Quinoa flour 2.8
0.1b,d,r 92
9b,d,e,k,l 58.7
1.5c,f,h,a,b 6.22
0.2a,b,c,k,m,g,h
Yeast e e 82.7
2.8 14.50
2.84

Control bread 1.7
0.07h,i,j,k,l 20.3
9f 63.8
2.2k,l,m,n,o,p,z 2.07
0.17b,s,w,x,z,a,b,c

Buckwheat bread 15 g/100 g 2.1
0.08e,h,m 33.4
3.5g 111.0
1.8r,u,k 5.42
0.17n,w,d
Buckwheat bread 30 g/100 g 2.65
0.1e,i,n 32.9
4h 150.8
2.2r,w,l 7.53
0.18n,x,3
Amaranth bread 15 g/100 g 1.73
0.09f,o 20.6
8i 73.7
1.6s,m,x 2.62
0.19o,p,t,z,4
Amaranth bread 30 g/100 g 2.61
0.04f,j 34.9
10j 114.1
3.0s,n,y 3.55
0.22o,r,u
Quinoa bread 15 g/100 g 1.88
0.07g,r 27.5
8.5k 71.0
3.1o,a 1.10
0.15a,t,a,g
Quinoa bread 30 g/100 g 2.54
0.1g,k 28.7
7l 76.5
1.3p,b 1.25
0.11b,p,r,s,u,b,h

e not analysed.
 J. Chlopicka et al. / LWT - Food Science and Technology 46 (2012) 548e555 551

breads e bread with addition of 15 g/100 g quinoa flour, 15 g/100 g drastic reductions of flavonoid concentrations (about 40%) in
amaranth flour and control bread made up of wheat flour had the comparison with groats manually dehulled. In the most severe
similar low total phenolic content: 1.88
0.07 mg/g dw, temperature treatment (the grains treated with steam (pressure
1.73
0.09 mg/g dw, 1.70
0.07 mg/g dw, respectively. Why 0.35 MPa, 164  C) for 20 min, followed by 50 min of treatment with
control bread had so low phenolic content is not clear for us and steam (0.4 MPa, 150  C) and final drying), the reduction of flavonoid
needs further investigation. content was 75%. It is not well understood which processes are
There were significant differences in respect to phenolic content responsible for the observed flavonoid losses. Dietrych-Szostak and
between all kinds of breads prepared either with addition of 15 g/ Oleszek (1999) concluded that such changes might be due to
100 g or 30 g/100 g of pseudocereals flour. The significant differ- flavonoid breakdown during heating and/or extraction of glyco-
ences in all cases between control bread and pseudocereals breads sides by the steam. In conclusion, the different thermal processing
also were observed, except for breads baked with 15 g/100 g could result in the decrease of flavonoids content, causing a drop in
addition of amaranth and quinoa flour, which had similar phenols antioxidative activities, particularly in buckwheat, but also, to less
content as wheat bread. The contents of phenols in breads were in extent, in amaranth and quinoa. Therefore in order to obtain
all cases lower than in respective flour. Based on works of health-promoting pseudocereals products, the processing condi-
Holtekjølen, Baevre, Rødbotten, Berg, and Knutsen (2008) and tion should be optimised to keep the loss of constituents as low as
Leenhardt et al. (2006) it is known that antioxidant active possible. Further evaluations in this field are required.
compounds presented in flours might be damaged or degraded as
a consequence of the heat/thermal process during baking. 3.3. Total antioxidant activities of flour and breads
However, losses of antioxidants during dough mixing and kneading
were also observed. Antioxidant activity of breads could be modi- Two methods were used to test the antioxidant activity of flours
fied by active oxidative enzymes presented in ingredients of and breads. First one (FRAP method) is based on determination of
compounds used in breads production, or oxidized by ambient ferric-tripyridyltriazine complex reducing capacity of the studied
oxygen. The addition of water will initiate enzyme activities, while extracts. Among polyphenols the greatest antioxidant efficacies in
a substantial incorporation of oxygen occurs during the initial this test were shown for quercetin, tannic acid, caffeic acid and
dough mixing and the remoulding into smaller pieces. Second gallic acid, while catechin and resveratrol had the lowest ones
obvious reason for lower antioxidant activity in breads as compared (Pulido, Bravo, & Saura-Calixto, 2000). Because of metal ion
with pseudocereal flours in current study was the fact that pseu- involvement in analytical reaction, FRAP method is fast, sensitive
docereal flours accounted for only 15e30 g for 100 g of all breads and spans relatively wide range of antioxidant substrates. However,
ingredients. the results obtained using FRAP method expressed the corre-
The phenols in control wheat bread were on similar level as sponding concentrations of electron-donating antioxidants, and
shown by Holtekjølen et al. (2008). Additionally, in accordance with the compounds that act by radical quenching, e.g. thiol antioxidants
our results, Alvarez-Jubete, Auty et al. (2009), Alvarez-Jubete, (such as glutathione) and carotenoids cannot be determined by this
Wijngaard et al. (2009) who investigated bread baked with 50 g/ assay. On the other hand, DPPH method is based on the evaluation
100 g addition of buckwheat, amaranth and quinoa flour, also

of the reducing ability of antioxidants toward DPPH , which is stable
observed the highest content of phenols in buckwheat bread. nitrogen radical, possessing an odd electron. The colour of its
However, in our investigation the level of phenolic compounds in solution fades rapidly when it encounters radical scavengers. In this
breads with amaranth flour ranged 1.73e2.61 mg/g and overlapped case, steric accessibility is a major determinant of the analytical
with phenolic compounds content in quinoa breads (1.88e2.54 mg/ reaction. Thus, this assay is adequate mainly for reactive small
g), while Alvarez-Jubete, Auty et al. (2009), Alvarez-Jubete, molecules that have good access to the radical site, and is less
Wijngaard et al. (2009) observed significantly lower phenolic sensitive for bigger molecules. The most effective antioxidants

concentration in amaranth bread than in quinoa bread. scavenging DPPH are gallic acid, tannic acid, ascorbic acid, and
quercetin.
3.2. Flavonoids contents of flour and breads These two methods (FRAP and DPPH) are complementary to
some degree. Both methods are recommended as easy, speedy,
The content of total flavonoids in flour was about 2e4 fold reproducible, and inexpensive for measuring the antioxidant
higher when compared to breads in all cases. All these differences activity in food extracts.
were statistically significant. Buckwheat flour was a better source of
flavonoid compounds than quinoa, wheat or amaranth flour 3.3.1. FRAP
(Table 2). Our results concerning total flavonoids in breads made The total antioxidant activities (TAA) determined by FRAP of
with addition of pseudocereals’ flour did not show such differences, flour and breads were expressed as mg of Trolox per 100 g dry
i.e. they did not depend on dosage of pseudocereals’ flour. The weight (Table 2). TAA of flour extracts, which were used to bake
lowest flavonoid level was presented in control bread. breads, was highest in case of buckwheat flour. TAA of wheat flour
According to the presented results the buckwheat breads was on the level of 158.3
3.0 mg Trolox/100 g dw. The antioxidant
showed a much higher loss of flavonoids than amaranth or quinoa. activity of quinoa and amaranth flours were the lowest two. All
Similarly, Zhang, Chen, Li, Pei, and Liang (2010) observed that total differences in antioxidant activities of evaluated flours were
flavonoids in buckwheat flour were decreased significantly after significant.
thermal processing such as roasting pressured steam-heating and The antioxidant activity in breads was highest for bread baked
microwave heating, while total phenolics were not influenced with 30 g/100 g addition of buckwheat flour, followed by the same
considerable. Sensoy et al. (2006) indicated that roasting (200  C dose of amaranth flour, and then bread with 15 g/100 g addition of
for 10 min) slightly reduced antioxidant activity of buckwheat flour. buckwheat flour. Remaining breads with amaranth and quinoa
Dietrych-Szostak and Oleszek (1999) noticed that even short flour had the similar lower antioxidant activity. The control bread
temperature treatment of buckwheat grain (heating for 10 min to had the lowest TAA. There were significant differences between
150  C) caused a significant decrease (20% of total) in flavonoid total antioxidant activity of all kinds of breads prepared with
concentration. Prolonged temperature treatment (heating process addition of either 15 g/100 g or 30 g/100 g pseudocereals flour, as
prolonged to either 1 h and 10 min or 2 h and 10 min) resulted in well as between control bread and pseudocereals breads in almost
552 J. Chlopicka et al. / LWT - Food Science and Technology 46 (2012) 548e555

all cases. TAA of wheat and buckwheat flours were significantly (i.e. between ferric-reducing capacity and nitrogen radical scav-
higher than in breads made from these flours. On the contrary, enging), but similar one in breads; (2) despite substantial loss of
opposed effects were found in TAA of flour and breads made with phenolics content during baking processes, still significant
addition of amaranth and quinoa seeds. In comparison with liter- contribution to antioxidant power and scavenging ability in
ature data (Alvarez-Jubete, Auty et al. 2009; Alvarez-Jubete, breads comprised phenolic compounds; (3) baking processes
Wijngaard et al. 2009) about antioxidant activity of pseudocereals reduced the variability and quantity of antioxidants in flour, but
flour, our results obtained by FRAP method were lower for those remaining in breads are equally well determined by both
amaranth, quinoa and buckwheat flour. Compared to the few data methods (FRAP and DPPH); therefore it would be interesting to
on FRAP in bread made with addition of buckwheat flour and evaluate more closely how much in common do these species
quinoa flour, available in the literature (Alvarez-Jubete, Auty et al. bear in respect to such features like their solubility and phase of
2009; Alvarez-Jubete, Wijngaard et al. 2009), our FRAP activity localization, reaction kinetics, and also physiologic aspects of
are in essential agreement. their varying reactivity.

3.3.2. DPPH 3.4. Sensory evaluation

The total antioxidant activities (TAA) determined by DPPH of
flours and breads were expressed as mmol of Trolox per 1 kg dry The results of hedonic scale analysis of colour, odour and
weight (Table 2). Total antioxidant activity (DPPH) was highest in consistency were shown in Table 3. Organoleptic evaluation
buckwheat flour, followed by quinoa flour and then wheat flour. (colour, odour, consistency) of breads revealed that in a ten-point
Amaranth flour had the lowest total antioxidant activity. The hedonic scale, all sensory results were in the range of 4e8 indi-
differences between buckwheat and quinoa flours and wheat and cating that these breads were moderately acceptable. The best
amaranth flours were statistically significant. colour of bread was observed in case of bread with 30 g/100 g
For breads, the total antioxidant activity in bread with addition addition of buckwheat flour, in contrast to bread with 15 g/100 g
of 30 g/100 g buckwheat flour was higher than that in bread with addition of quinoa flour, which had the worst colour of all exam-
addition of 15 g/100 g buckwheat flour. Similar results were ob- ined breads. The similar observation about buckwheat was pre-
tained in buckwheat bread by Alvarez-Jubete, Auty et al. (2009), sented by Lin et al. (2009), who suggested that buckwheat
Alvarez-Jubete, Wijngaard et al. (2009). Breads with the addition of improved colour of wheat breads because it contains more phenolic
amaranth flour (independently on dose of flour) and control bread compounds which could inhibit the browning processes during
had higher total antioxidant activity than bread with addition of baking.
30 g/100 g quinoa flour. Bread with addition of 15 g/100 g quinoa Bread with 30 g/100 g addition of buckwheat flour had the best
flour had the lowest antioxidant activity. There were significant odour of all breads, and bread with addition of 15 g/100 g of
differences between total antioxidant activities of all kinds of amaranth flour had the worst one. The results suggested that
breads depending on percentage of flour, except breads made up of substituting 15 g/100 g of amaranth flour in the bread formula
quinoa flour. The significant differences for total antioxidant would not be advantages for bread acceptability. The best influence
activity (DPPH) between control bread and investigated pseudo- of odour on evaluated breads had a buckwheat flour, and this effect
cereals breads were also observed in all cases. The DPPH scavenging was previously observed by Lin et al. (2009).
activity of wheat bread was similar to data of Martinez-Villaluenga The consistency of bread was the best in case of bread with the
et al. (2009). addition of 15 g/100 g of buckwheat. Consistency of control bread
TAA (DPPH) of all kinds of flours was higher than in breads made and amaranth bread were of the same quality. The worst consis-
up of these flours (significant differences were in case of wheat and tency was observed in bread with 15 g/100 g addition of quinoa
quinoa flours and breads). flour.
The decreasing levels of TP, FRAP, DPPH in breads in comparison The statistically significant difference in colour was found only
to the flours used as an additional ingredient were revealed in all between bread with 30 g/100 g addition of buckwheat flour and
cases. These results suggest that the antioxidants, which were bread with 15 g/100 g addition of quinoa flour (p < 0.01). In case of
present in cereals, can be modified during thermal processes. Other odour the significant difference was only between bread with 15 g/
authors made similar observations (Holtekjølen et al., 2008; 100 g addition of amaranth flour and bread with 30 g/100 g addi-
Martinez-Villaluenga et al. 2009; Moore, Luther, Cheng, & Yu, tion of buckwheat flour (p < 0.01). No statistically significant
2009). differences were found in consistency of examinated breads, but in
There were positive and significant correlations between total respect to colour and consistency breads with quinoa flour had the
phenolic compounds and FRAP of breads (r ¼ 0.710; p < 0.0001), worse scores of organoleptic analysis. Breads with buckwheat and
total phenolic compounds and DPPH of breads (r ¼ 0.455; amaranth flour had a good valuation of their features, and these
p < 0.01), FRAP and DPPH of breads (r ¼ 0.924; p < 0.001). results suggest that these kinds of breads were pretty acceptable for
However, there was no correlation between FRAP and DPPH of consumers.
flours. All these results suggest, that: (1) there was different trend The effects of the addition of pseudocereals flour into wheat
in antioxidant capacity between FRAP and DPPH in flours flour on the taste of breads was shown in Figs. 1e3.

Table 3
The ratings of control bread and breads with addition of pseudocereals flour (median values and 95% confidence intervals for medians; the scorecard was developed with 10-
point category scale (disliked ¼ 0; extremely liked ¼ 10), number of testers n ¼ 31; results marked with the same letter in upper index within each row differ significantly,
p < 0.01).

Attributes Bread

Control Buckwheat Buckwheat Amaranth Amaranth Quinoa Quinoa

bread bread 15 g/100 g bread 30 g/100 g bread 15 g/100 g bread 30 g/100 g bread 15 g/100 g bread 30 g/100 g
Colour 8 (7e9) 7 (5e7) 8 (7e8)a 6 (5e7) 7 (6e8) 5 (4e6)a 6 (6e7)
Odour 7 (6e9) 6 (4e7) 8 (7e8)b 5 (4e7)b 7.5 (5e7) 7.5 (5e7) 7 (6e8)
Consistency 7 (6e9) 7.5 (5e7) 7 (6e8) 7 (5e7) 7 (6e7) 4 (2e6) 7 (6e8)
 J. Chlopicka et al. / LWT - Food Science and Technology 46 (2012) 548e555 553

60
intresting

milk 50 tasty

40
delicate natural

30
difficult to
determine
20 strange taste

10

0 vapid
tart

not to eat
salty

bad taste
sweet

little pronounced
gummy

crusty

Fig. 1. Spider diagram of the sensory evaluation [%] of the different buckwheat breads and the comparison to the control bread. *Breads were tested 12 h after backing. % values in
the diagram were calculated as % of testers (n ¼ 31) who approved respective feature. Blue line e % of positive answers given by testers to control bread; green line e % of positive
answers given by testers to bread with addition of 15% buckwheat flour; yellow line e % of positive answers given by testers to bread with addition of 30% buckwheat flour.
(For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

60
intresting

milk 50 tasty

40
delicate natural

30
difficult to
determine
20 strange taste

10

0 vapid
tart

not to eat
salty

bad taste
sweet

little pronounced
gummy

crusty

Fig. 2. Spider diagram of the sensory evaluation [%] of the different amaranth breads and the comparison to the control bread. *Breads were tested 12 h after backing. % values in the
diagram were calculated as % of testers (n ¼ 31) who approved respective feature. Blue line e % of positive answers given by testers to control bread; green line e % of positive
answers given by testers to bread with addition of 15% amaranth flour; red line e % of positive answers given by testers to bread with addition of 30% amaranth flour.
(For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)
554 J. Chlopicka et al. / LWT - Food Science and Technology 46 (2012) 548e555

70
intresting
60
milk tasty

50
delicate natural
40

difficult to 30
determine
strange taste
20

10

0 vapid
tart

not to eat
salty

bad taste
sweet

little pronounced
gummy

crusty

Fig. 3. Spider diagram of the sensory evaluation [%] of the different quinoa breads and the comparison to the control bread. *Breads were tested 12 h after backing. % values in the
diagram were calculated as % of testers (n ¼ 31) who approved respective feature Blue line e % of positive answers given by testers to control bread; brown line e % of positive
answers given by testers to bread with addition of 15% quinoa flour; yellow line e % of positive answers given by testers to bread with addition of 30% quinoa flour. (For inter-
pretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

The influence of buckwheat flour is shown in Fig. 1. The char- addition of buckwheat flour into bread can improve not only
acteristics of buckwheat bread taste indicate that bread with antioxidant but also sensory properties of bread.
addition of 30 g/100 g of buckwheat flour had an interesting and
natural taste and was crusty as more than 40% of testers declared 4. Conclusions
such opinion. Control bread and bread with 15 g/100 g of buck-
wheat flour were not crusty and testers declared gummy taste. The addition of buckwheat flour to wheat flour improved more
Results suggest that buckwheat flour (30 g/100 g) improved the effectively antioxidant status of bread than other studied pseudo-
sensory value of bread and reduced attributes (i.e. gummy), which cereals, amaranth and quinoa. Buckwheat bread had a highest
could decrease value of bread. Bread with 30 g/100 g buckwheat content of phenolic compounds.
flour had a highest scores of sensory profile such as in buckwheat The sensory profile results suggest that the buckwheat bread
bread examination by Wronkowska et al. (2008). has more positive sensory value than amaranth and quinoa breads.
The influence of amaranth flour is shown in Fig. 2. In case of Results showed also that addition of buckwheat flour up to 15 g/
amaranth bread it was noticed that it had some negative opinions 100 g or 30 g/100 g levels to wheat flour improved satisfactory
(about 10e30% testers) stating that this bread was “not to eat”, bread properties and attributes such as colour, odour and taste.
gummy, and had a bad, strange or difficult to determine taste. Only These results indicate that consumers may willingly choose buck-
30% testers declared an interesting taste of amaranth bread. wheat breads.
Therefore, the addition of amaranth flour to wheat bread does not
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