Food Chemistry 125 (2011) 164–170

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Food Chemistry
journal homepage: www.elsevier.com/locate/foodchem

Feasibility of use of buckwheat flour as an ingredient in ginger nut

biscuit formulation
Bojana Filipčev ⇑, Olivera Šimurina, Marijana Sakač, Ivana Sedej, Pavle Jovanov, Mladenka Pestorić,
Marija Bodroža-Solarov
Institute for Food Technology, University of Novi Sad, Bul. cara Lazara 1, 21000 Novi Sad, Serbia

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

Article history: Ginger nut biscuits are popular traditional biscuits which contain honey. In order to improve their nutri-
Received 28 April 2010 tive profile and functionality, standard formulation based on wheat flour was substituted with buckwheat
Received in revised form 18 June 2010 flour at three levels (30%, 40%, 50%). The quality attributes (chemical composition, microelements, bioac-
Accepted 24 August 2010
tive compounds (total polyphenols, rutin, quercetin), antioxidative potential, textural and sensory prop-
erties) were compared to control biscuit (based on wheat flour) and biscuits substituted with rye at the
same levels, since rye is used as an ingredient which improves the quality of ginger nut biscuits. Substi-
Keywords:
tution with buckwheat flour resulted in a significant increase (p < 0.05) in protein, Zn, total polyphenols,
Biscuits
Buckwheat
antioxidative and chelating activity. Total dietary fibre, Cu, Mn and Fe contents also increased but did not
Rutin exceed those in rye substituted biscuits. Consumption of 100 g of enriched biscuits could contribute to
Quercetin reaching 18–22% (buckwheat) and 16–17% (rye) of the estimated daily average total polyphenols intake,
Antioxidative potential respectively. With the contents of rutin and quercetin at 3.96–6.57 mg/100 g d.b. and 0.087–0.214 mg/
Quality 100 g d.b., respectively, buckwheat substituted biscuits may significantly contribute to total dietary flavo-
noid intake and meet the demands of preventive nutrition. Buckwheat enriched biscuits were rated higher
regarding sensory attributes, softness and fracturability as compared to control but lower compared to rye
supplemented biscuits.
Ó 2010 Elsevier Ltd. All rights reserved.

1. Introduction flavonoids, phenolic acids, tannins, phytosterols, fagopyritols and

Common buckwheat is a pseudocereal that used to be an impor-
tant part of human diet but it has been neglected especially since
the prevalence of the Western dietary pattern. However, buck-
wheat has recently received a renewed interest on the basis of
its nutritional composition and its feasibility for natural, organic
cultivation.
Buckwheat is a rich source of starch, proteins, fibres, antioxi-
dants and minerals (especially iron, manganese, zinc and sele-
nium) (Christa & Soral-Śmietana, 2008; Krkošková & Mrázová,
2005). Some of its components were found to exhibit outstanding
health-promoting properties: its proteins were shown to have cho-
lesterol lowering properties (Christa & Soral-Śmietana, 2008; Kreft
& Germ, 2008; Krkošková & Mrázová, 2005); it is suitable for celiac
patients as its prolamins do not exert immunological responses
like those in cereals (Krkošková & Mrázová, 2005); substantial
amounts of resistant starch may contribute to the lowering of gly-
caemic index of food (Christa & Soral-Śmietana, 2008; Kreft &
Germ, 2008). Among other compounds with prophylactic value,
⇑ Corresponding author. Tel.: +381 21 450 3778; fax: +381 21 450 725.
E-mail address: bojana.filipcev@fins.uns.ac.rs (B. Filipčev).
lignans were identified. The main buckwheat flavonoid is rutin,
used for medicinal purposes to strengthen capillary walls, reduce
high blood pressure and increase microcirculation. It also improves
C vitamin absorption and eye health. Fagopyritols are derivatives
of D-chiro-inositol, a special carbohydrate compound, which play
an important role in insulin signal transduction and were found
to be deficient in patients suffering from Type II Diabetes and Poly-
cystic Ovary Syndrome. Prophylactic properties of these com-
pounds can be associated to their high antioxidant activity.
Flavonoids and phenolics in buckwheat seeds manifested antilipo-
peroxidative potential in the studies of Oomah and Mazza (1996),
Watanabe, Oshita, and Tsushida (1997) and Watanabe (1998).
Buckwheat seeds demonstrated higher lipid stability protection
factor than oats and barley (Holasova et al., 2002). In the recent
study of Takahama, Tanaka, and Hirota (2009) it was reported that
that the ingestion of dough prepared from buckwheat reduced the
nitric acid present in stomach juices to nitric oxide which is impor-
tant since its presence at gastric level has been shown to exert sev-
eral important biological functions like antimicrobial activity,
improvement in gastric motility, increase in mucus production
and mucosal blood flow (Peri et al., 2005). In addition, this may
prevent the formation of toxic nitrosamines in the digestive tract.

0308-8146/$ - see front matter Ó 2010 Elsevier Ltd. All rights reserved.
doi:10.1016/j.foodchem.2010.08.055
 B. Filipčev et al. / Food Chemistry 125 (2011) 164–170 165

Consequently, buckwheat has been considered by many In the present research work, the inclusion of buckwheat flour
researchers as a highly valuable ingredient with tremendous po- at various levels to ginger nut biscuit formulation was investigated
tential in designing functional food products (Holasova et al., in order to assess the optimal formulation, characterise the
2002; Krkošková & Mrázová, 2005; Skrabanja et al., 2004; Hatcher, improvement in the nutritional value and biological functionality
You, Dexter, Campbell, & Izydorczyk, 2008; Kreft, Fabjan, & Yasum- and evaluate textural and sensory properties of the products. An-
oto, 2006; Wijngaard & Arendt, 2006). Traditionally, food uses of other objective of the paper was to compare buckwheat substi-
buckwheat include mainly home-made dishes like porridge, stews, tuted biscuits with those containing rye in the same proportion.
pilaffs, casseroles and pancakes in which buckwheat is used in the
form of groats (coarsely cracked hulled hydrothermally treated 2. Material and methods
grains), grits (finely ground groats) or flour. Wholemeal flour is
prepared by milling the whole buckwheat grain similarly to wheat. 2.1. Materials
Light flour is made by milling hulled buckwheat groats. Blends of
30% buckwheat and 70% wheat flour are commonly used in house- Dough samples were prepared from ingredients purchased from
holds for bread preparation in Slovenia (Kreft & Germ, 2008). In Ja- local suppliers: commercial soft wheat flour type 500) (moisture
pan and Italy, buckwheat flour mixed with wheat flour is used for content 14.03%, ash 0.53% dry basis), honey, vegetable fat, sugar,
preparation of noodles. NaHCO3, lecithin and spices (cinnamon and clove). Wholegrain
Today, several trends in food uses of buckwheat have been antic- rye flour (moisture content 9.37%, ash 1.61% d.b.) and wholegrain
ipated: its use in improving traditional foods, development of new buckwheat flour (moisture content 12.31%, ash 2.20% d.b.) were
functional foods, development of food additive from buckwheat obtained from a local healthy-food store. Rye flour had finer gran-
with special biological effects and better use of buckwheat by-prod- ulation (72.08% of particles were less than 150 lm) whereas buck-
ucts (Krkošková & Mrázová, 2005). Therefore, it is not odd that the wheat flour was coarse (48.89% of particles was larger than
number of new products containing buckwheat has been increasing 350 lm).
in the market along with the popularity of functional foods. In the
literature, divers reports can be found dealing with nutritional, 2.2. Ginger nut biscuit making procedure
functional and/or processing characterisation of buckwheat based
functional foods: wheat bread enriched with extracts obtained from Ingredients used for the preparation of ginger nut biscuits are
green buckwheat plants (Gawlik-Dziki, Dziki, Baraniak, & Rufa, listed in Table 1.
2009), wheat bread enriched with husked and unhusked buck- The biscuits were prepared according to a two-phase procedure.
wheat at 15% flour substitution level (Lin, Liu, Yu, Lin, & Mau, In the first phase, a mixture of honey, sugar and water was heated
2009) and soba noodles (Hatcher et al., 2008), etc. Alamprese, Casi- to 65 °C. In the second phase, after cooling to 40 °C, lecithin, flour
raghi, and Pagani (2007) successfully included buckwheat into glu- blends, NaHCO3 and spices were mixed using a kitchen mixer
ten-free pasta formulations. Kreft et al., (2006) broadened the equipped with a spiral hook. Water was added in amount to pro-
investigation to less common buckwheat products such as buck- duce dough with acceptable handling characteristics. Dough con-
wheat leaf flour, buckwheat beer and vinegar. sistency and stickiness were subjectively estimated by an
However, the use of buckwheat in so-called convenient food or experienced baker. Dough moisture ranged between 17–20% for
ready-to-eat food products such as biscuits, breakfast cereals and the control and rye substituted biscuits and 15.0–15.5% for buck-
other confectionary products seems less extensive. This is probably wheat biscuits. Dough samples were covered with a plastic foil
due to limited production caused by low crop yields and a small and left to rest over night at 15 °C. After that, the dough was rolled
market. Consequently, less literature data can be found on the to- on a pastry board to uniform thickness of 10 mm and cut into 6 cm
pic. Several attempts were made to include buckwheat flour into diameter rounds. Biscuits were baked at 170 °C for 10–15 min.
this category of products, such as in the trials of Śmietana et al.
(Śmietana, Fornal, Szpendowski, Soral-Śmietana, & Fornal, 1985; 2.3. Chemical analyses
Śmietana, Fornal, Szpendowski, Soral-Śmietana, & Fornal 1988)
where buckwheat was used as a component mixed with milk pro- Standard methods of analysis (AOAC, 2000) were used to deter-
teins and cereals. Fornal, Śmietana, Soral-Śmietana, Fornal, and mine crude protein (950.36), fat (935.38), reducing sugar (975.14),
Szpendowski (1985), Fornal, Śmietana, Soral-Śmietana, Fornal, total dietary fibre (958.29), and water contents (926.5). In the pro-
and Szpendowski (1987) investigated the extrusion process of mix- tein determination, a nitrogen-to-protein conversion factor of 6.25
tures containing buckwheat flour and characterised the obtained was used. Starch content was determined by hydrochloric acid dis-
products. Im, Huff, and Hsieh (2003) studied the effect of process- solution according to the ICC Standard (ICC Standard No. 123/1
ing conditions on the physical and chemical properties of cakes ob- 1994) using the specific optical rotation for wheat starch 182.7
tained by puffing and moulding buckwheat grits. grd ml/g dm1 in calculation.
Ginger nut biscuits are products with a long tradition. They are
a popular type of soft and delicious biscuits containing honey and
spices. The first gingerbread is thought to have been made by Cath- Table 1
Formulations of ginger nut biscuits.
olic monks in Europe for special holidays and festivals. Explorers
and warriors of the middle ages introduced spices from Asia and Ingredients, g Control Buckwheat substituted Rye substituted
the near East to Europe which were later adopted by cake-maker ginger nut biscuit ginger nut biscuit
artisans who started the production of spicy sweet honey cakes. Wheat flour 100 70 60 50 70 60 50
Traditional recipes for ginger nut biscuits vary across Europe Buckwheat flour 0 30 40 50 0
Rye flour 0 0 30 40 50
depending on the spice blend, type and quantity of honey used
Honey 50 50 50
and applied technology. In the 19th century, the production of gin- Sugar 10 10 10
ger nut biscuits was mechanized and approached the phase of Vegetable fat 10 10 10
industrial production. In both industrial and artisan practise, NaHCO3 2.1 2.1 2.1
wheat flour is frequently substituted with rye flour in doses up Spice blend 2 2 2
Lecithin 1 1 1
to 20% which contribute to better moisture retention and pro-
Water 13.5 12.5 12 11.3 18 20 21.2
longed freshness of the product.
166 B. Filipčev et al. / Food Chemistry 125 (2011) 164–170
All analyses were performed in duplicate. Three composite sam-
ples of each biscuit type were analysed. The composite samples
were prepared by homogenisation of six individual biscuit
samples.
2.4. Determination of mineral composition
Mineral composition was determined using a Varian Spectra AA
10 (Varian Techtron Pty Limited, Mulgvare Victoria, Australia,
1989) atomic absorption spectrophotometer equipped with a
background correction (D2-lamp). The sample preparation con-
sisted of a dry ashing procedure at 450 °C as described in Pavlović,
Kevrešan, Kelemen-Mašić, and Mandić (2001).
Limits of detection (LOD) and quantification (LOQ) calculated on
the basis of three and ten times the standard deviation of the mean
blank determinations from 15 measurements were 0.015 mg/100 g
and 0.02 mg/100 g for Fe; 0.0125 and 0.02 mg/100 g for Cu;
0.005 mg/100 g and 0.01 mg/100 g for Mn; 0.0025 mg/100 g and
0.005 mg/100 g for Zn, respectively.
The accuracy of the procedure applied for determination of the
minerals in ginger nut biscuits was validated by analysis of certi-
fied reference material NIST SRM 1567a Wheat flour (National
Institute of Standard and Technology, Gaithersburg, MD, USA).
The mean recovery percentages for the studied elements obtained
from eight replications confirmed the accuracy of the procedure:
Fe (101.6 ± 2.51), Mn (95.9 ± 3.23), Cu (105.3 ± 4.47) and Zn
(101.7 ± 3.10).
2.5. Preparation of ethanolic extracts
Ethanolic extracts were prepared by suspending 5 g of pow-
dered biscuits in 50 ml of ethanol/water (80/20, v/v). After 1-h
shaking, the suspension was left overnight at ambient tempera-
ture. Following extraction, the sample was filtered through filter
paper (Whatman, Grade 4 Chr, UK), and the procedure was re-
peated twice with 50 ml of solvent. The resulting supernatants
were combined and dried by vacuum-evaporator. The dried extract
was redissolved in ethanol/water (80/20, v/v) to 10 ml volume and
used to determine total phenolics and antioxidative potential.
2.6. Determination of total phenolics
Total phenolics analysis was based on the Folin–Ciocalteu
method (Singleton, Orthofer, & Lamuela-Raventos, 1999). Extracts
(0.1 ml) were diluted with pure water (7.9 ml). Folin–Ciocalteu’s
reagent (0.5 ml) and Na2CO3 solution (1.5 ml, concentration 20 g/
100 ml) were added and mixed thoroughly. The mixture was al-
lowed to stand for 120 min with intermittent shaking, and the
absorbance at 750 nm was measured (Jenway, 6405 UV/Vis). A gal-
lic acid calibration curve (c = 97.107  A + 0.0101, R2 = 0.9984) was
plotted on the basis of seven calibration points within the range of
1.00–6.00 lg/ml of gallic acid in the reaction mixture. The total
phenolics content of ginger nut biscuits were expressed as gallic
acid equivalents (GAE) (mg GAE/100 g on dry mass basis).
2.7. Determination of rutin and quercetin
One gram of biscuit was extracted with 20 ml methanol/water
(90/10, v/v) at room temperature during 24 h, ultrasonicated for
10 min followed by filtration through a 0.45 lm pore size nylon fil-
ter (Rotilabo-Spritzenfilter 13 mm, Roth, Karlsruhe, Germany) be-
fore injection into the HPLC system.
HPLC analysis was performed by using a liquid chromatograph
(Agilent 1200 series), equipped with a diode array detector (DAD),
on an Agilent, Eclipse XDB-C18, 1.8 lm, 4.6  50 mm column, at a
flow-rate of 1.000 ml/min. Based on the previously used method
(Cetkovic et al., 2008), new chromatographic conditions were
developed. The solvent linear gradient programme was created
by varying the proportion of solvent A (methanol) to solvent B
(1% formic acid in water (v/v)) as follows: initial 10% A; 0–
10 min, 10–25% A; 10–20 min, 25–60% A; 20–30 min, 60–70% A.
The run time and post-run time were 45 and 10 min, respectively.
The column was operated at 30 °C. The injected volume of samples
and standards was 5 ll and it was carried out automatically, using
an autosampler. The spectra were acquired in the range 210–
400 nm and chromatograms plotted at 280 and 330 nm with a ref-
erence wavelength set at 550/100 nm.
Rutin and quercetin in a sample extract were identified by
matching the retention time and their spectral characteristics
against those of the standards. The purity of the peaks was deter-
mined to ensure the identification. The external standard method
was a technique used for quantification.
2.8. Performance of the chromatographic method
Stock solutions of rutin and quercetin standards were prepared
at a concentration of 1.0 mg/ml in methanol. The solutions were di-
luted with 1% formic acid to obtain a series of dilutions in the range
of 0.005–34 lg/ml in methanol:1% formic acid in water (1:1, v/v)
for external standard calibration and linearity check at 280 and
330 nm for rutin and quercetin, respectively. The limits of detection
(LOD) and quantification (LOQ) under the given chromatographic
conditions, determined at a signal-to-noise (S/N) ratio of 3 and
10, respectively, were: for rutin LOD = 0.009 lg/ml, LOQ =
0.039 lg/ml; and for quercertin LOD = 0.039 lg/ml, LOQ =
0.131 lg/ml. The linearity range for rutin was obtained within the
concentration range 0.009–17 lg/ml (y = 3546.39x  0.0849,
R2 = 0.9999), while obtained linearity range for quercetin was be-
tween 0.05–20 lg/ml (y = 10056x + 0.293, R2 = 0.9997). The reten-
tion time and peak area repeatability was checked by injecting
the mixture of rutin and quercetin, at the concentration of
0.010 mg/ml for each standard, into the HPLC system over six runs.
The within- and inter-day relative standard deviation (RSD) of the
retention times was less than 0.2% and for the obtained peak area
less than 3%, for both tested standards.
2.9. Determination of antioxidative potential
2.9.1. Antioxidative activity (AOA) in b-carotene/linoleic acid model
system
AOA by b-carotene bleaching method as described in detail by
Moure et al. (2000) was used. Degradation rate of extracts was cal-
culated as proposed by Al-Saikhan, Howard, and Miller (1995).
AOA was expressed as % inhibition relative to the control using
an Eq. (1):
Degradation rate of control  Degradation rate of sample
AOA ¼
Degradation rate of control
 100
ð1Þ
IC50 value (mg/ml) was defined as the concentration of extract
at which 50% inhibition of free radical activity is observed under
the experimental conditions. It was derived by interpolation from
linear regression analysis.
2.9.2. Reducing power
Reducing power of the extracts was determined in accordance
with the method of Oyaizu (1986). Extracts (0.50–20.0 mg/ml)
were mixed with equal volumes of 0.2 M phosphate buffer (pH
6.6) and 1% potassium ferricyanide and then incubated at 50 °C
for 20 min. Afterwards, 2.5 ml 10% trichloroacetic acid was added.
 B. Filipčev et al. / Food Chemistry 125 (2011) 164–170 167

After centrifugation at 650g for 10 min, 0.5 ml FeCl3 and 2.5 ml of
distiled water was added to 2.5 ml of supernatant and the absor-
bance was measured at 700 nm (Jenway, 6405 UV–Vis). Reducing
power was expressed as IC50 value i.e. an extract concentration
at which the absorbance of reaction mixture reaches 0.5.
2.9.3. DPPH radical scavenging activity
The method was adopted from Hatano, Kagawa, Yasuhara, and
Okuda (1988). 0.1 ml of extract (0.50–20.0 mg/ml), 1.0 ml of DPPH_
(90 lmol/l) and 2.9 ml methanol were vigorously mixed and left to
stand in the dark for 60 min. Then, absorbance at 517 nm was
measured.
Results were expressed as IC50 value or the concentration of ex-
tract required to scavenge 50% of the initial amount of DPPH_.
2.10. Fe2+ chelation activity
Chelating activity was measured according to the method of
Decker & Welch, 1990). It was calculated from the absorbances
of the control and samples using an Eq. (2):
Chelating activity ð%Þ
Absorbance of sample at 562 nm  100
¼ 100   100
Absorbance of control at 562 nm
Results were interpolated and expressed as IC50 value or the ex-
tract concentration which chelates 50% of present Fe2+ ions under
the given conditions.
2.11. Water activity measurement
at 5 g. From the force–time plot, two parameters were derived: the
area under the curve represented the biscuit hardness and the lin-
ear distance was an indication of fracturability. Eight measure-
ments per each biscuit type were made.
2.13. Physical characteristics and sensory evaluation
Biscuits were evaluated for different parameters. Six, randomly
chosen biscuits were taken to measure weight, height and width
(diameter). Spread was calculated from the ratio of width and
height.
Quality characteristics of biscuits included the estimation of
several parameters: shape, appearance of upper surface, appear-
ance of lower surface, fracture, structure, chewiness, and flavour.
The scores for each parameter ranged from 1 to 5. Each parameter
was assigned a different factor of significance: 0.3 for shape, 0.5 for
upper surface, 0.4 for lower surface, 0.4 for fracture, 0.6 for struc-
ture, 0.8 for chewiness and 1.0 for flavour. Total score was obtained
by summing the scores for each parameter, previously multiplied
with the corresponding factor. A panel of seven judges was en-
gaged in the biscuit scoring.
ð2Þ
2.14. Statistical analysis
Data were evaluated by analysis of variance (ANOVA), general
linear models of the Statistica 7.1 statistical software (StatSoft
Inc., Tulsa, Oklahoma). Means were compared using the Tukey’s
test at the 0.05 level of probability.

Water activity was measured using an Aw-meter, model Testo
650 (Testo, Germany) with accuracy ± 0.001. Tests were conducted
24 h post-bake on a composite sample prepared from six biscuits
broken into small pieces at 21 °C. Parallel measurements were
done for each sample.
2.12. Textural analysis
Textural analysis of ginger nut biscuits was performed by punc-
turing using a TA.HDplus Texture Analyzer (Stable Micro Systems,
England, UK) according to embedded project ‘‘Biscuit penetration”
B1S1-P2.PRJ. The analyzer was set to measure force in compres-
sion. A 2 mm cylinder probe and a 5 kg load cell were used. Each
biscuit was precisely centred on the platform with a holed plate
and the probe was lowered to puncture the sample at a travel dis-
tance of 20 mm and test-speed 0.5 mm/s. The trigger force was set
3. Results and discussion
3.1. Proximate composition
Proximate composition of ginger nut biscuits is outlined in Ta-
ble 2. According to the literature data, the minimal required mois-
ture content of ginger nut biscuits is 7% (Gavrilović, 2003). All the
samples tested met this criterion. Below this value, the biscuits are
prone to excessive drying and hardening. The buckwheat supple-
mented biscuits had significantly lower moisture contents of all
samples, assuming as the consequence of low moisture absorption
of rather coarsely milled wholegrain buckwheat flour. Isolated
buckwheat starch was reported to have higher water-binding
capacity than wheat and corn starch (Wijngaard & Arendt, 2006)
but it seems that the moisture absorption behaviour of buckwheat
flour is much influenced by its granulation.

Table 2
Chemical composition of buckwheat and rye supplemented ginger nut biscuits (given on dry basis).

Parameters Control Buckwheat Rye

30% 40% 50% 30% 40% 50%
Moisture content, g/100 g 9.47 ± 0.12b 8.94 ± 0.08a 8.56 ± 0.08a 8.70 ± 0.09a 11.78 ± 0.11c 11.30 ± 0.12c 11.72 ± 0.13c
Protein content, g/100 g 7.22 ± 0.15a 7.73 ± 0.13b 7.84 ± 0.11b,c 8.12 ± 0.17c 7.14 ± 0.12a 7.00 ± 0.11a 7.16 ± 0.13a
Fat content, g/100 g 8.12 ± 0.15a 8.43 ± 0.26a 8.45 ± 0.20a 8.59 ± 0.17a 8.51 ± 0.13a 8.16 ± 0.26a 8.37 ± 0.09a
Starch content, g/100 g 45.80 ± 1.84b 43.64 ± 1.28a,b 43.77 ± 1.19a,b 43.21 ± 1.25a,b 44.08 ± 1.36a,b 42.03 ± 1.44a,b 41.05 ± 1.26a
Total reducing sugar content, g/ 29.37 ± 1.15b,c 25.33 ± 1.18a 25.90 ± 1.10a 27.29 ± 0.97a,b 29.73 ± 0.88b,c 32.11 ± 1.18c 31.17 ± 0.94c
100 g
Total dietary fibres, g/100 g 3.87 ± 0.13a 5.49 ± 0.22b 5.37 ± 0.53b 7.61 ± 0.34c 6.47 ± 0.55b,c 8.48 ± 0.34c,d 8.87 ± 0.76d
Zn, mg/100 g 0.34 ± 0.06a 0.78 ± 0.10b,c 1.00 ± 0.12c,d 1.16 ± 0.09d 0.60 ± 0.04b 0.70 ± 0.05b 0.81 ± 0.08b,c
Cu, mg/100 g 0.07 ± 0.03a 0.15 ± 0.02a,b 0.16 ± 0.03a,b 0.22 ± 0.05b 0.12 ± 0.03a 0.14 ± 0.03a,b 0.16 ± 0.04a,b
Mn, mg/100 g 0.61 ± 0.02a 0.88 ± 0.04b 0.88 ± 0.06b 0.93 ± 0.03b 0.95 ± 0.10b 1.05 ± 0.12b,c 1.20 ± 0.09c
Fe, mg/100 g 0.57 ± 0.07a 1.39 ± 0.13b 1.66 ± 0.16b 1.72 ± 0.15b 1.58 ± 0.10b 1.99 ± 0.17b,c 2.38 ± 0.21c
Rutin content, mg/100 g Not detecteda 3.96 ± 0.19b 5.24 ± 0.03c 6.57 ± 0.21d Not detecteda Not detecteda Not detecteda
Quercetin content, mg/100 g Not detecteda 0.087 ± 0.0005b 0.143 ± 0.0169c 0.214 ± 0.0110d Not detecteda Not detecteda Not detecteda
Total phenolics, mg GAE/100 g 157.06 ± 3.77a 196.35 ± 0.90d,e 202.58 ± 2.10e 238.92 ± 1.81f 177.51 ± 1.95c 167.90 ± 0.33b 191.45 ± 4.76d

a, b, c, d,. . . Different superscripts within the same raw represent significant differences between the results (p < 0.05).
168 B. Filipčev et al. / Food Chemistry 125 (2011) 164–170
Partial replacement of wheat flour with buckwheat flour con-
siderably improved the protein, total dietary fibre and mineral con-
tent, especially that of Zn, Mn and Fe, expectedly. Likewise, the
replacement with rye flour increased the content of fibres, Zn,
Mn, Fe and reducing sugars. The addition of buckwheat or rye flour
did not affect the fat content. The starch content showed a decreas-
ing trend with increasing supplementation levels but it was signif-
icant only in the case of 50% rye supplemented biscuit in
comparison to the control.
3.2. Content of polyphenols
As expected, supplemented biscuits exhibited significantly
higher total polyphenols contents (Table 2) as compared to control
which increased with increasing supplementation levels of both
buckwheat and rye. Within biscuits with 30% and 40% buckwheat
and 50% rye, there was no significant difference in the content of
total phenolics. Biscuit containing 50% buckwheat was the highest
in the polyphenols content (238.92 mg/100 g dry basis (d.b.)). Ru-
tin and quercetin were not detected in the control and rye supple-
mented biscuits for apparent reasons. Increased levels of
buckwheat flour increased rutin (3.96–6.57 mg/100 g d.b.) and
quercetin (0.09–0.21 mg/100 g d.b.) contents (Table 2). This is sev-
eral times higher than the contents of rutin and quercetin reported
for the buckwheat bread (15% buckwheat flour on replacement ba-
sis) which ranged between 0.90–1.75 mg/100 g d.b. and 0.03–
0.04 mg/100 g d.b., respectively (Lin et al., 2009).
Rutin (quercetin-3-O-rutinoside) and quercetin (the precursor
of rutin) are the major bio-functional compounds of buckwheat
belonging to the category of flavanol glycosides. Rutin is often de-
noted as vitamin P1 whereas quercetin has been addressed as the
main flavanol in human diet. Various prophylactic effects of buck-
wheat, including increased antioxidant, anti-inflammatory and
anti-tumour effects have been associated with the activity of rutin
and quercetin. Beneficial effects of quercetin range from high anti-
oxidant activity and anti-inflammatory capacity to miscellaneous
effects like anti-fibrotic, anti-bacterial, anti-hypertensive, anti-ath-
erogenic and anti-proliferative (Boots, Haenen, & Bast, 2008). The
doses of rutin which demonstrated therapeutical effects have been
reported to range between 180 and 350 mg (Schilcher, Patz, &
Schimmel, 1990). The consumption of 100 g of ginger nut biscuits
supplemented with buckwheat at the tested doses would only cov-
er approximately 1–3% of the therapeutical dose. Although official
dietary reference intake for rutin has not yet been established, ad-
vice suggesting the intake of 10–25 mg/day of rutin for adults and
5–10 mg/day for children can be found elsewhere (Anon., 2006,
2010). In this view, 100 g of biscuit containing 50% of buckwheat
could contribute to appreciable 24–60% of the proposed daily dose
for adults.
Data on average daily dietary intakes of polyphenols are rather
scanty due to a number of limitations. Estimations showed that the
total intake of polyphenols reaches approximately 1000 mg/day in
case of subjects who have a habit to daily consume several portions
of fruits, phenolic acid-rich beverages, and vegetables (Manach,
Table 3
Antioxidative potential profile (IC50, mg/ml) of buckwheat and rye supplemented ginger nut biscuits.
Control Buckwheat
30%
Antioxidative activity (AOA) 32.51 ± 0.67d 26.71 ± 0.29b
Reducing activity 29.36 ± 0.05c 28.46 ± 0.72c
DPPH_ scavenging activity 23.06 ± 0.87e 10.79 ± 0.26c,d
Chelating activity 11.24 ± 0.06a 11.84 ± 0.10a
a, b, c, d, e Different superscripts within the same raw represent significant differences between the results (p < 0.05).
Scalbert, Morand, Remesy, & Jimenez 2004; Scalbert & Williamson,
2000). Consumption of 100 g of ginger nut biscuits made with
buckwheat and rye could contribute to reaching 18–22% and
16–17% of the estimated average intake, respectively.
The intake of flavonoids as a special class of polyphenolic com-
pounds, was reported to vary widely: from 20 to 1000 mg/day
(Kühnau, 1976). More recent studies revealed that less than
20 mg/day quercetin is administered by food (Mullie, Clarys, Derie-
maeker, & Hebbelinck, 2007). Very lower intake of quercetin,
3.3 mg/day was estimated in the Finnish population (Knekt, Järvi-
nen, & Reunanen, 1996; Knekt et al., 2002). The intake of rutin and
quercetin would be at 3.61–6.00 mg/day and 0.08–0.19 mg/day,
respectively, by consumption of 100 g of buckwheat biscuits. All
in all, products enhanced with buckwheat may significantly con-
tribute to total dietary flavonoid intake and hence, facilitate to
meeting the demands of preventive nutrition.
3.3. Antioxidative potential of ginger nut biscuits
Profiling of antioxidant potential of ginger nut biscuits was as-
sessed here by applying four different antioxidant assays (Table 3).
The results expressed as IC50 values (mg/ml) for better comparison.
Lower IC50 values are indication of higher antioxidant potential.
Antioxidative activity of biscuits measured by the inhibition of
bleaching of b-carotene followed the order: control = 30%
rye < 40% = 50% rye < 30% = 40% buckwheat < 50% buckwheat.
There was a strong correlation between AOA IC50 and total pheno-
lics content (r = 0.90) suggesting that phenolic compounds pre-
sented in all produced biscuits mainly contributed to their
overall antioxidant properties.
The ability to scavenge DPPH_ by the extracts of rye and buck-
wheat supplemented biscuits was significantly higher (p < 0.05)
than the ability of an extract obtained using the control biscuit.
Strong scavenging activity on DPPH_ of buckwheat enriched biscuit
extracts was partly due to the presence of rutin which possesses a
strong ability to scavenge DPPH_ (Yang, Guo, & Yuan, 2008).
There is a correlation (r = 0.72) between the results obtained in
the reducing activity assay and DPPH assay which can be explained
by the same principles of these methods. One of the important
mechanisms to protect against oxidative damage and lipid peroxi-
dation is to chelate metal ions. Distinct differences in chelating
activity on Fe2+ were observed between the buckwheat and rye
supplemented biscuits (Table 3). It is likely that the enrichment
of biscuits with buckwheat flour contributed to metal chelating
activity on Fe2+ since rutin has been evidenced as a potent metal
chelator (Yang et al., 2008). Moreover, there are experimental data
supporting that wheat is also a potent Fe2+ chelator (personal cor-
respondence) which can explain good chelating activity of the con-
trol sample.
The obtained results indicated a benefit of using both buck-
wheat and rye flours in order to achieve better functionality of pro-
duced biscuits. Buckwheat enriched biscuits were characterised
with higher antioxidant potential in comparison to rye enriched
biscuits.
Rye
40% 50% 30% 40% 50%
25.37 ± 0.10b 23.83 ± 0.38a 31.10 ± 0.92d 29.06 ± 0.68c 28.66 ± 0.36c
28.00 ± 0.28b,c 26.2 ± 0.1a 28.02 ± 1.17b,c 26.61 ± 0.17a,b 25.26 ± 0.09a
9.66 ± 1.28b,c 5.25 ± 1.12a 12.30 ± 0.46d 12.05 ± 0.1c,d 8.10 ± 0.94b
11.35 ± 0.70a 11.21 ± 0.62a 18.52 ± 0.36c 17.98 ± 0.47c 15.92 ± 0.16b
 B. Filipčev et al. / Food Chemistry 125 (2011) 164–170 169

Table 4
Physical characteristics and texture of ginger nut biscuits supplemented with buckwheat and rye flour.

Ginger nut biscuit Dose,% Weight, g Height, mm Diameter, Spread ratio Density, g/ Water Hardness, g s Fracturability, g s
type mm cm3 activity
Control 0 28.52 ± 1.29a 17.33 ± ± 0.22a 63.23 ± 0.61c 3.65 ± 0.06c 0.51 ± 0.02a,b 0.50 ± 0.00a 6724.14 ± 736.36c 1166.12 ± 164.31c
Rye flour 30 30.22 ± 1.39a 20.12 ± 0.61b 62.73 ± 0.76b,c 3.12 ± 0.08a 0.49 ± 0.03a 0.58 ± 0.01c 1922.17 ± 351.13b 383.60 ± 43.63b
40 30.37 ± 0.89a 20.71 ± 0.52b 62.42 ± 0.49b,c 3.02 ± 0.07a 0.49 ± 0.01a,b 0.60 ± 0.02c 2289.36 ± 210.69b 450.46 ± 31.86b
50 28.87 ± 1.42a 20.53 ± 1.16b 62.13 ± 0.62b 3.03 ± 0.20a 0.46 ± 0.03a 0.59 ± 0.01c 1596.23 ± 206.47b 349.30 ± 24.79b
Buckwheat flour 30 29.60 ± 0.87a 17.87 ± 0.77a 61.57 ± 0.60b 3.46 ± 0.14b,c 0.55 ± 0.01b,c 0.55 ± 0.02b 3925.26 ± 275.82a 789.65 ± 57.70a
40 29.62 ± 0.96a 17.88 ± 0.54a 61.66 ± 0.43b 3.45 ± 0.11b,c 0.56 ± 0.01c 0.53 ± 0.02a,b 3473.08 ± 497.97a 663.59 ± 81.80a
50 29.92 ± 1.17a 17.72 ± 0.46a 60.56 ± 0.51a 3.42 ± 0.10b 0.59 ± 0.02c 0.54 ± 0.01b 4413.35 ± 366.37a 795.78 ± 68.51a

a, b, c Different superscripts within the same raw represent significant differences between the results (p < 0.05).

3.4. Physical characteristics of ginger nut biscuits

Data on the physical and textural characteristics of ginger nut

biscuits are presented in Table 4. There were no significant differ-
ences in the weight of biscuits. The thickness was significantly in-
creased by the addition of rye flour at all levels whereas buckwheat
flour did not significantly affect this parameter. The diameter of
cookies was reduced significantly except in the case of biscuits
made with 30% and 40% rye flour. The biscuits with buckwheat
(40%, 50%) had significantly higher density as compared to the con-
trol and rye supplemented biscuits.
Supplemented biscuits exhibited a decreased spread which ran-
ged from 3.65 (control) to 3.02 (40% rye). Only samples made with
30% and 40% buckwheat flour did not significantly differentiate
from the control. According to Gurjal, Mhta, Samra, & Goyal,
2003), two factors affect the spread ratio: expansion of dough by
leavening and gravitational flow. Dough flow mostly depends on
its viscosity: the higher the viscosity the lower the spread rate.
Fuhr (1962) suggested that any ingredient that absorbs water dur-
ing dough mixing will decrease the spread ratio. Rye flour was a
better absorbent of water than buckwheat flour presumably be-
cause buckwheat flour contained more coarse particles which held
less water in comparison to the fine particles of rye flour.
Water activity, a measure of food dryness and susceptibility of a
product to microbial spoilage, increased in the ginger nut biscuit
formulations with rye and buckwheat. Although less water was
needed to make homogeneous dough in the case of buckwheat bis-
cuits, their aw was significantly higher than that of the control.
Granular structure of buckwheat biscuits may have affected this
increase. The higher water activity values found in these supple-
mented cookies (up to 0.60 for biscuit with 40% rye flour) are not
likely to increase their susceptibility to microbial growth (Fontana,
2000).
The biscuits differentiate in hardness and fracturability. Two
groups, significantly softer from the control, could be observed.
The softest were the biscuits supplemented with rye flour whereas
biscuits made with buckwheat flour were softer from the control
but harder from those made with rye. A similar grouping was ob-
served regarding fracturability. The most fracturable biscuit was
the control sample. The least fracturable were the biscuits made Fig. 1. Sensory evaluation and total sensory score of ginger nut biscuits supple-
mented with buckwheat and rye flour.
with rye.

3.5. Sensory evaluation of ginger nut biscuits
As seen in Fig. 1, the highest sensory scores were given to the
biscuits supplemented with rye, of which the best were the formu-
lations at 30% and 40% level (18.3 points). The biscuits made with
buckwheat flour were also scored significantly higher than the
control with the highest score given to the formulation at 40% level
(17.4 points). In general, these groups of biscuits differentiated in
chewiness and structure. The supplemented biscuits were softer
than the control and therefore rated higher for chewiness. The bis-
cuits with added buckwheat had a drier mouthfeel than those
made with rye. The structure of biscuits made with the addition
of rye was more porous. Buckwheat flour contributed to a porous
but mainly granular structure whereas the control sample was
compact. It is important to note that the characteristic aroma of
buckwheat was completely masked even at the 50% supplementa-
tion level.
4. Conclusion
Buckwheat has been scientifically proven to exert a wide range
of bioactivities with beneficial health effects. However, its propor-
170 B. Filipčev et al. / Food Chemistry 125 (2011) 164–170
tion in the product formulation and processing methods influence
the content of health-promoting compounds in the end-product.
Therefore, it is useful to characterise the product in order to assess
the content and functionality of its active compounds. Moreover,
data on the levels of active compounds in foods is helpful in quan-
tifying their dietary intake.
Buckwheat can be incorporated into ginger nut biscuit up to
50% as it provides a sensorially acceptable product with enhanced
nutritional and bio-functional properties. The sensory analysis
indicated the product with 40% buckwheat flour was the best
(within buckwheat supplemented ones) but we are of the opinion
that a futher improvement of the structure of 50% buckwheat bis-
cuit could be achieved by using buckwheat flour with finer
granulation.
Biscuit supplementation with buckwheat flour resulted in a sig-
nificant increase (p < 0.05) in protein, Zn, total polyphenols, antiox-
idative and chelating activity. Total dietary fibre, Cu, Mn and Fe
contents also increased but did not exceed those in the rye substi-
tuted biscuits. Consumption of 100 g of buckwheat enriched bis-
cuits could contribute to reaching 18–22% of the estimated
daily average total polyphenols intake and could significantly con-
tribute to total dietary flavonoid intake and meet the demands of
preventive nutrition.
The inclusion of buckwheat in the formulations of convenient
food is beneficial for consumers as it may contribute to more var-
iant and balanced diet.
Acknowledgement
The authors acknowledge the financial support of the Ministry
of Science and Development of the Republic of Serbia as a part of
the Project 20068.
References
Al-Saikhan, M. S., Howard, L. R., & Miller, J. C. Jr, (1995). Antioxidant activity and
total phenolics in different genotypes of potato (Solanum tuberosum L.). Journal
of Food Science, 60, 341–347.
Alamprese, C., Casiraghi, E., & Pagani, M. A. (2007). Development of gluten-free fresh
egg pasta analogues containing buckwheat. European Food Research Technology,
225, 205–213.
Anon, (2010) Acu-cell nutrition: Bioflavonoids rutin-hesperidin (http://acu-
cell.com/bio.html, accessed on Feb. 1, 2010).
Anon, (2006). Nature’s Corner newsletter vol. 3, 29
www.naturescorner.com/news8_5_06.html, accessed on Feb. 1, 2010).
AOAC (2000). In: W. Horwitz (Ed.), Official methods of analysis of AOAC
international, 17th ed. Maryland: USA.
Boots, A. W., Haenen, R. M. M. G., & Bast, A. (2008). From antioxidant to
nutraceutical. European Journal of Pharmacology, 585, 325–337.
Ćetković, G., Čanadanović-Brunet, J., Ðilas, S., Savatović, S., Mandić, A., & Tumbas, V.
(2008). Assessment of polyphenolic content and in vitro antiradical
characteristics of apple pomace. Food Chemistry, 109, 340–347.
Christa, K., & Soral-Śmietana, M. (2008). Buckwheat grains and buckwheat
products-nutritional and prophylactic value of their components – a review.
Czech Journal of Food Science, 26(3), 153–162.
Decker, E. A., & Welch, B. (1990). Role of ferritin as a lipid oxidation catalyst in
muscle food. Journal of Agricultural and Food Chemistry, 38, 674–677.
Fontana, A. J. (2000). Understanding the importance of water activity in food. Cereal
Foods World, 45, 7–10.
Fornal, Ł., Śmietana, Z., Soral-Śmietana, M., Fornal, J., & Szpendowski, J. (1985).
Products extruded from buckwheat flour and its mixtures with milk proteins. II.
Chemical characteristics and physico-chemical properties of proteins and
starch products extruded buckwheat flour and its mixtures with milk
proteins. Acta Alimenatria Polica, 11, 397–411.
Fornal, Ł., Śmietana, Z., Soral-Śmietana, M., Fornal, J., & Szpendowski, J. (1987).
Chemical characteristics and physico-chemical properties of the extruded
mixtures of cereal starches. Starch/Stärke, 39, 75–78.
Fuhr, F. R. (1962). Cookie spread: Its effects on production and quality. Bakers’
Digest, 36, 56–60.
Gavrilović, M. (2003). Proizvodnja medenjaka i srodnih proizvoda (Production of
ginger nut biscuits and similar products). In F. Stanković (Ed.), Tehnologija
konditorskih proizvoda (Technology of confectionery products) (2nd ed.) (pp. 552–
559). Novi Sad: University of Novi Sad.
Gawlik-Dziki, U., Dziki, D., Baraniak, B., & Rufa, L. (2009). The effect of simulated
digestion in vitro on bioactivity of wheat bread with Tartary buckwheat flavones
addition. LWT-Food Science and Technology, 42, 137–143.
Gurjal, H. S., Mhta, S., Samra, I. S., & Goyal, P. (2003). Effect of wheat bran, coarse
wheat flour and rice flour on the instrumental texture of cookies. International
Journal of Food Properties, 6(2), 329–340.
Hatano, T., Kagawa, H., Yasuhara, T., & Okuda, T. (1988). Two new flavonoids and
other constituents in licorice root: Their relative astringency and radical
scavenging effects. Chemical and Pharmaceutical Bulletin, 36, 2090–2097.
Hatcher, D. W., You, S., Dexter, J. E., Campbell, C., & Izydorczyk, M. S. (2008).
Evaluation of the performance of flours from cross- and self-polinating
Canadian common buckwheat (Fagopyrum esculentum Möench) cultivars in
soba noodles. Food Chemistry, 107, 722–731.
Holasova, M., Fiedlerova, V., Smrcinova, H., Orsak, M., Lachman, J., & Vavreinova, S.
(2002). Buckwheat-the source of antioxidant activity in functional foods. Food
Research International, 35, 207–211.
Im, J.-S., Huff, H. E., & Hsieh, F.-H. (2003). Effects of processing conditions on the
physical and chemical properties of buckwheat grit cakes. Journal of Agricultural
and Food Chemistry, 51, 659–666.
Knekt, P., Järvinen, R., & Reunanen, A. (1996). Flavonoid intake and coronary
mortality in Finland: A cohort study. British Medical Journal, 312, 478–481.
Knekt, P., Kumpulaien, J., Järvinen, R., Rissanen, H., Heliövaara, M., Reunanen,
Hakulinen., et al. (2002). Flavonoid intake and risk of chronic disease. American
Journal of Clinical Nutrition, 76, 560–568.
Kreft, I., Fabjan, N., & Yasumoto, K. (2006). Rutin content in buckwheat (Fagopyrum
esculentum Möench) food materials and products. Food Chemistry, 98, 508–512.
Kreft, I., & Germ, M. (2008). Organically grown buckwheat as a healthy food and a
source of natural antioxidants. Agronomski glasnik, 4, 397–406.
Krkošková, B., & Mrázová, Z. (2005). Prophylactic components of buckwheat. Food
Research International, 38, 561–568.
Kühnau, J. (1976). The flavonoids: a class of semi-essential food components: Their
role in human nutrition. World Review Nutrition Diet, 24, 117–191.
Lin, L.-Y., Liu, H.-M., Yu, Y.-W., Lin, S.-D., & Mau, J.-L. (2009). Quality and antioxidant
property of buckwheat enhanced wheat bread. Food Chemistry, 112, 987–991.
Manach, C., Scalbert, A., Morand, C., Remesy, C., & Jimenez, L. (2004). Polyphenols,
food sources and bioavailability. American Journal of Clinical Nutrition, 79,
727–747.
Moure, A., Cruz, J. M., Franco, D., Domínguez, J. M., Sineiro, J., Domínguez, H., et al.
(2001). Natural antioxidants from residual sources. Food Chemistry, 72,
145–171.
Mullie, P., Clarys, P., Deriemaeker, P., & Hebbelinck, M. (2007). Estimation of daily
human intake of food flavonoids. Plant Foods in Human Nutrition, 62, 93–98.
Oomah, B. D., & Mazza, G. (1996). Flavonoids and antioxidative activities in
buckwheat. Journal of Agricultural and Food Chemistry, 44, 1746–1750.
Oyaizu, M. (1986). Studies on products of browing reaction–antioxidant activities of
products of browing reaction prepared from glucoamine. Japanese Journal of
Nutrition, 44, 307–315.
Pavlović, A., Kevrešan, Ž., Kelemen-Mašić, Ð., & Mandić, A. (2001). Micro and toxic
elements in skin, meat and gills of grass carp (Ctenopharyngodon idella). In
Proceedings of the 8th symposium on analytical and environmental problems (pp.
117–122). Szeged, Hungary: SZAB.
Peri, L., Pietraforte, D., Scorza, G., Napolitano, A., Fogliano, V., & Minetti, M. (2005).
Apples increase nitric oxide production by human saliva at the acidic pH of the
stomach: a new biological function for polyphenols with a catechol. Free Radical
Biology and Medicine, 43, 1233–1242.
Scalbert, A. & Williamson, G. (2000). Dietary intake and bioavailability of
polyphenols. The Journal of Nutrition, 130(8S), 2073S–2085S.
(http://
Schilcher, H., Patz, B., & Schimmel, K. (1990). Klinische Studie mit einem
Phytopharmacon zur Behandlung von Mikrozirkulationsstorungen.
Ärztzeitschrift fur Naturheilverfahren, 31, 819–826.
Singleton, V. L., Orthofer, R., & Lamuela-Raventos, R. M. (1999). Analysis of total
phenols and other oxidation substrates and antioxidants by means of Folin-
Ciocalteu reagent. Methods in Enzymology, 299, 152–178.
Skrabanja, V., Kreft, I., Golob, T., Modic, M., Ikeda, S., Ikeda, K., et al. (2004). Nutrient
content in buckwheat milling fractions. Cereal Chemistry, 81, 172–176.
Śmietana, Z., Fornal, Ł., Szpendowski, J., Soral-Śmietana, M., & Fornal, J. (1985).
Products extruded from buckwheat flour and its mixtures with milk proteins. I.
Technological aspects of extrusions and their relationship with the expansion
and porosity of structure. Acta Alimentaria Polonica, 3, 275–284.
Śmietana, Z., Fornal, Ł., Szpendowski, J., Soral-Śmietana, M., & Fornal, J. (1988).
Utilization of milk proteins and cereal starches to obtain co-extrudates.
Nahrung, 32, 545–551.
Takahama, U., Tanaka, M., & Hirota, S. (2009). Proanthocyanidins in buckwheat flour
can reduce salivary to nitric oxide in the stomach. Plant Foods in Human
Nutrition, published online 15 December 2009. doi:10.1007/s11130-009-0144-
x.
Yang, J., Guo, J., & Yuan, J. (2008). In vitro antioxidant properties of rutin. LWT-Food
Science and Technology, 41, 1060–1066.
Watanabe, M. (1998). Catechins as antioxidants from buckwheat (Fagopyrum
esculentum Möench) groats. Journal of Agricultural and Food Chemistry, 46,
839–845.
Watanabe, M., Oshita, Y., & Tsushida, T. (1997). Antioxidant compounds from
buckwheat (Fagopyrum esculentum Möench) hulls. Journal of Agricultural and
Food Chemistry, 45, 1039–1044.
Wijngaard, H. H., & Arendt, E. K. (2006). Buckwheat. Cereal Chemistry, 83(4),
391–401.