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Health quality and nutritional value of rye bread produced on a small and large
scale in poland

Article  in  Italian Journal of Food Science · October 2013

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 OPINION

HEALTH QUALITY AND NUTRITIONAL VALUE

OF RYE BREAD PRODUCED ON A SMALL
AND LARGE SCALE IN POLAND

M. RADZYMINSKA*, B. GARBOWSKA and D. JAKUBOWSKA

Chair of Commodity Science and Food Research, University of Warmia and Mazury in Olsztyn
*Corresponding author: [email protected]

Abstract

This study is a small part of a project aimed at performing comprehensive evaluation of the
quality of local raw materials and products in the region of Warmia and Mazury in Poland. This
study evaluates selected determinants of the health quality and nutritional value of rye bread in
relation to origin (small and large scale production). The analytical procedure included determi-
nation of the content of acrylamide, organochlorine pesticides and polychlorinated biphenyls, n-
methylcarbamates, ochratoxin A, acidity, moisture, protein, glucose, fat, salt, Cu, Mn, Fe, Zn, Ca,
and Mg. The quality of bread produced on a small scale, compared to that produced in large bak-
eries, differed only in terms of some determinants. Rye bread from local producers contained sta-
tistically significantly less (P ≤ 0.05) acrylamide and statistically significantly more copper, man-
ganese and zinc.

- Keywords: traditional products, acrylamide, organochlorine pesticides, polychlorinated biphenyls,

n-methylcarbamates, ochratoxin A, Cu, Mn, Fe, Zn, Ca and Mg -

126  Ital. J. Food Sci., vol. 25 - 2013

 INTRODUCTION the characterization of even the typical senso-
ry traits of the traditional products.
Now that globalization of the food economy Bread made from whole and brown rye flours
has made the market of food products predom- is a major component of diet in Northern and
inated by mass products whose quality param- Eastern European countries (MICHALSKA et al.,
eters frequently fail to meet consumers’ expec- 2008). As bread is a staple food to many popula-
tations, local agri-food initiatives, proposed as tions, its quality is of outmost importance. Being
sustainable alternatives, are gaining impor - a component of an everyday diet, bread should
tance both for small-scale producers and en- have high nutritional quality (DEWETTINCK et
vironmentally-aware consumers (SELFA and al., 2008). It has been shown to be a source of
QAZI, 2005; O’HARA and STAGL, 2001; HINRICKS, nutrients, especially carbohydrates, fiber, pro-
2000). The level of consumers’ trust in food and teins and some minerals (magnesium, phospho-
perception of risks vary, but purchasing de- rus, iron) (KOPEĆ et al., 2011). As reported by
cisions are increasingly often associated with MICHALSKA et al. (2008), rye products based on
seeking safe food products (YEUNG and MOR- wholemeal rye flour may be considered to be nu-
RIS, 2001; VERBEKE et al., 2007; ANGULO and tritious and healthy food.
GIL, 2007) with well-known and tested places The chemical composition, nutritional value
of origin (ESPEJEL et al., 2007; DU PLESSIS and and sensory quality of bread depend on the type
DU RAND 2011). The region of origin is begin- and extract of rye flour used for its production,
ning to play an important role in decisions of on the technological process, water content and
food products purchase (LOBB and MAZZOCCHI, technological additives (KIHLBERG et al., 2004)
2007). Those from rural areas are perceived as which impart the product specific functional
products of high quality (HENCHION and MCI- properties (JONES and JEW, 2007). Modification
NTYRE, 2000). of the recipe by the addition of some ingredients,
”Traditional food” refers to products made for example, dietary fiber and sour dough, may
from specific raw materials, and/or with a reci- affect the nutritional value of the finished prod-
pe known for a long time, and/or with a specif- uct (LOPEZ et al. 2003; POUTANEN et al., 2009;
ic process (CAYOT, 2007). The current EU legal TUCKER et al., 2010).
regulations (COUNCIL REGULATION No 2081/92; An increasing number of products are appear-
No 2082/92; No 510/2006; No 509/2006) ena- ing on the market with names given by produc-
ble the protection of original agricultural prod- ers and implying that they were “home-made”.
ucts and food characteristic of the place of or- By information provided on packages, the pro-
igin and traditional methods of production. ducers suggest that the food was manufactured
These include: Protected Designation of Origin with traditional methods, according to tradition-
(PDO), Protected Geographical Indication (PGI) al recipes.
and Traditional Speciality Guaranteed (TSG). There are only scarce data on local food. Infor-
Quality of food, resulting from the method of its mation about products manufactured on a small
production and processing, as well as the raw scale, from local raw materials and with tradi-
materials used, should be the most important tional methods, are frequently only uncorrobo-
attribute of regional food and the base its pos- rated press reports. The aim of this study was to
itive image should be built on. The term ‘‘tra- compare: 1) health quality (acrylamide, Σ DDT, γ
ditional’’ is, however, not adequately defined. HCH, PCB, N-methylcarbamates, ochratoxin A,
Moreover, it is often intentionally or uninten- and 2) nutritional value (protein, glucose, Cu,
tionally misused. Therefore, the important is- Mn, Fe, Zn, Ca and Mg) of rye bread produced
sues of registration and standardization of tra- by micro-companies and larger producers.
ditional foods arise in order for these products
(1) to be protected against imitations, (2) to be
of high quality and (3) to conform to contempo- EXPERIMENTAL PROCEDURES
rary rules of appropriate and safe production. A
way to ensure authenticity and high quality of Samples
traditional food products is to establish criteria
for their registration that will thereafter deter- During the first stage, an inventory was per-
mine standards for their commercial produc- formed of bread produced by agri-tourist farms
tion (TRICHOPOULOU et al., 2007). Reproduc- and micro-companies in the bakery industry,
tion of traditional food, especially today, in the located in the region of Warmia and Mazury
era of globalisation, entails a number of ques- in Poland (Fig. 1). Seventy-five personal inter-
tions about the possibility of obtaining compo- views with representatives of the entities under
nents, reconstructing a recipe and a process, study were conducted. The questionnaire con-
the nutritional value, specificity or typicality tained three groups of questions: 1) concerning
of the products. CAYOT (2007) has pointed out the types of products made, 2) focusing on raw
that the industrialization of food production, materials and methods of production, 3) aimed
European laws on food safety and even the de- at providing a qualitative description of prod-
velopment of innovative products necessitate ucts made from local raw materials. 20% of the

Ital. J. Food Sci., vol. 25 - 2013  127

 ses were carried out using Shimadzu LC-20AD
series for HPLC equipped with UV-Vis 190-
600 nm diode array detector (DAD, Shimad-
zu) set at 200 nm, a vacuum degasser, binary
pumps, thermostated autosampler and tempera-
ture-controlled column oven. The column used
was C18, 4.0 mm guard column and Synergi 4
Hydro-RP 80 A 250 x 4.6 mm LC column from
Phenomenex (Torrance, CA); mobile phase: iso-
cratic mixture of 5 mM heptasulfonic acid; flow
rate 1.0 mL/min; injection volume: 100 µL, col-
umn temperature 25°C, autosampler tempera-
ture 4°C; UV-Vis detector set at 200 nm. Acryl-
amide elutes near 8-9 min. Total time between
injections was 30 min. Peak identification was
based on the retention time by comparison of
the ratio of UV spectra with that of a standard
commercial compound. The acrylamide stan-
dard was obtained from Sigma-Aldrich Chemi-
Fig. 1 - Map of rye bread collection cal Company (USA). Stock standards were pre-
pared in 80% methanol. The stock solution of
the standard was 20, 10, 5, 2, 1, 0.5, 0.25, 0.10,
0.05 and 0.01 µm/mL. All standard solutions
companies under study were found to use only were stored at 4°C in glass light-resistant bot-
flour purchased from local, regional producers. tles. The linearity of the standard curve was ex-
3% of the companies used materials originating pressed in terms of the correlation coefficient
from their own production. The place of origin (R2), the plot of the integrated peak area (% of
of materials was an important factor, taken into UV spectra) against standard concentration. A
account in purchasing decisions for 45% of the linear regression equation was found with lin-
companies. The analyzed products were those earity (R2=0.9996, n=6) over the range of 0.01 -
with higher-than-average quality, resulting from 20 µg/mL. This was used to determined acryl-
traditional methods of production and materials amide concentration in food samples. Repeat-
of local origin used in the production process. ability and precision of the method were deter-
Of all the products under study, samples of mined by repetitive analyses of the content of
rye bread from 22 manufacturers were collected acrylamide in 10 one-batch samples of rye bread
(the samples were labelled as traditional food). and calculating the RSD (relative standard de-
Moreover, bread labelled as “wholemeal bread” viation). The RSD (n=10) of one-batch samples
was bought on the Warmia and Mazury market was 4.7% which is a satisfactory level in food
(Poland) from 8 large-scale manufacturers (the quality control. The mean recovery was 105%.
samples were labelled as conventional). Organochlorine pesticides (DDT and γ HCH)
and polychlorinated biphenyls (PCBs) were ex-
Health quality tracted from the samples with petroleum ether
and acetone together with fat. The identification
Health quality of rye bread was determined and quantitative determination of the analyzed
based on selected indicators: acrylamide, Σ DDT compounds were carried out with gas chroma-
(1.1.1. - trichloro-2.2-bis(4-chlorophenyl)eth- tography using a PYE Unicam 4600 apparatus
ane - as a total sum of DDT + DDE +DDD), γ with an EC detector.
HCH (γ-hexachlorocyclohexane), polychlorinat- N-methylcarbamates were determined with
ed biphenyls (PCBs), N-methylcarbamates and the HPLC method (DIONEX CORPORATION 2007;
ochratoxin A. ISO 1998). The method of sample preparation in-
The content of acrylamide in the samples was volves a salting-out extraction step with acetoni-
determined with the RP-HPLC method (WENZL trile, NaCl, and MgSO4; and a SPE clean-up step
et al., 2003; ZHANG et al., 2005). The 4.0-g sam- using primary secondary amine (PSA) resin to
ple after acrylamide extraction using 80% meth- extract carbamates and remove interfering sub-
anol in water and de-fatting using hexane was stances from the samples. 5 g of the sample were
cleaned up by adding acetonitrile (20 mL) and put into a tube and then 5 ml of acetonitrile were
Carrez I (500 µL) and Carrez II (500 µL). The al- added. After 1 min of vortexing, 2 g of MgSO4 and
iquot was centrifuged for 10 min at 4,500 g at 0.5g NaCl were added and the sample was mixed
4°C and the clear supernatant was cleaned up for 1 min using vortex. Then, it was centrifuged
with Oasis HLB SPE cartridges purchased from for 10 min (rpm 3,000) and 1 mL of supernatant
Waters (Milford, MA). The final sample was fil- was pipetted into 1.5 mL and 100 mg MgSO4 and
tered through a membrane filter (0.45 µm) and 50 mg PSA were added. Afterwards, the sample
analyzed with the RP-HPLC method. The analy- was vortexed for 1 min and centrifuged for 5 min

128  Ital. J. Food Sci., vol. 25 - 2013

(10,000 rpm). Then, 100 µL of the supernatant es using petroleum benzine and in gravimetric
were mixed with 900 µL of distilled water and an- determination of the amount of fat collected in
alyzed with the HPLC. The following separation the receiver of the Soxhlet apparatus.
conditions were applied: Shimadzu LC-20AD liq- Determination of calcium and magnesium
uid chromatograph; column: guard column: C18 contents were conducted according to the meth-
Acclaim Carbamate 3.0 x 10 mm, 3 µm, analytical od described by WHITESIDE and MIMER (1984).
column: Acclaim Carbamate 3.0 x 150 mm, 3 µm; The samples were mineralized in a mixture of ni-
column temperature: 50°C; mobile phase: meth- tric and perchloric acids (03:01). Mineralization
anol-water in gradient; flow rate 0.9 mL/min; in- was conducted in an electric aluminium heat-
jection volume: 50 µL; detector fluorescent (exci- ing block with temperature programming (VELP
tation: 330 nm, emission: 465 nm). Peak identi- DK 20, manufactured by VELP Scientifica, Ita-
fication was based on the retention time by com- ly). Contents of Ca and Mg in the mineral resi-
parison with the standard. due were determined using flame atomic absorp-
Ochratoxin A was determined with the HPLC tion spectroscopy (acetylene-air flame). Meas-
method with bicarbonate clean up according to urements were carried out with the use of atom-
ISO 15141-2 (ISO 1998). Extraction of ochratox- ic absorption spectrometer Unicam 939 Solar -
in A was made using SPE-cartridges purchased Great Britain, equipped with an Optimus data
from Waters (Milford, MA). After extraction, the station, background correction (deuterium dis-
level of OTA was determined with HPLC using charge lamp) and an appropriate cathode lamp.
Shimadzu LC-20AD series for HPLC Chromato- A 10% aqueous lanthanum chloride solution, in
graph with reversed phase column (250 x 4.6 amounts ensuring a final concentration of La3+
mm, 5 µm C18). A mixture of acetonitrile, water at 1%, was added to all measured solutions to
and glacial acetic acid at the ratio of 49·5:49·5:1 determine Ca content. Standards of calcium and
and the flow rate of 1 mL/min was used in the magnesium (1 mg/cm3), diluted with 0.1 M so-
analysis. A fluorimetric detector was used as lution of HNO3, were prepared on the basis of
well (excitation wavelength 330 nm, emission BDH standards (Germany).
wavelength 460 nm). Qualitative identification In addition, bread was subjected to the evalu-
was performed by comparing the retention times ation of its physicochemical parameters includ-
of the standard with that of the sample under ing: acidity, salt content and moisture content.
study and quantitative determination was per- Determination of potential acidity was speci-
formed by comparing the areas of peaks of the fied by titration of the examined solution with a
standards with those of the sample under study. base of a known concentration, in the presence
of phenolphthalein as an indicator.
Nutritional value Determination of salt content in bread was
performed with the Mohr’s method. The princi-
Nutritional value of rye bread was determined ple of the method consists in titrating an aque-
based on selected indicators: protein, glucose, ous extract of the pulp with silver nitrate, in the
fat, Cu, Mn, Fe, Zn, Ca, and Mg. presence of potassium chromate as an indicator.
Protein content in bread was determined ac- Determination of bread moisture was per-
cording to Kjeldahl’s method. This method con- formed with the application of a drying meth-
sists in mineralization (burning) of the examined od. Such determination consists in specifying
organic substance in concentrated sulfuric acid, the loss of water from a sample while drying it
at boiling temperature. The examined substance at 130°C for 60 minutes.
oxidizes to carbon dioxide and water, and the ni-
trogen it contains gets separated in the form of Data analysis
ammonia. About 2 g of the sample were miner-
alized and distilled. The mineralization and dis- Data obtained were analyzed statistically with
tillation process was performed in the FOSS ap- the use of basic statistics, i.e. mean and stand-
paratus. After distillation, the samples were ti- ard deviation and relative standard deviation.
trated with 0.1 M of hydrochloric acid. Protein Differences between health quality and nutri-
content in bread was expressed in grams per tional value categories of bread (traditional and
100 g of the product. conventional) were determined with the use of
Glucose content was determined using the one-way analysis of variance (ANOVA). The sig-
Lane-Eynon method. The principle for determin- nificance of differences was tested at a signifi-
ing sugars according to this method is copper re- cance level of 0.05.
duction in Fehling I and II (because of sugars pre-
sent in the examined sample) and complete con-
version of bivalent into monovalent copper (by ti- RESULTS AND DISCUSSION
trating with a standard solution of saccharose)
in the presence of the indicator (methylene blue). Average concentrations of selected indicators
Determination of fat content was carried out of bread quality in all examined samples in rela-
using the Weibull-Stoldt method. The principle of tion to their origin (traditional and convention-
the method consists in extracting fat substanc- al) are presented in Table 1.

Ital. J. Food Sci., vol. 25 - 2013  129

Table 1 - Average values of selected determinants in the bread samples under study.

Bread
Traditional n= 66 Conventional n= 40
×− ± SD RSD ×− ± SD RSD
Acrylamide μg/kg 119.62± 37.11b 31.02 163.35±26.22a 16.05
Σ DDT mg/kg of fat 0.032±0.034 106.25 0.028±0.029 103.57
γ HCH mg/kg of fat 0.001±0.001 100 0.002±0.001 50
PCB mg/kg of fat 0.007±0.003 42.86 0.005±0.004 80
N-methylcarbamate mg/kg No found No found
Ochratoxin A mg/kg No found No found
Acidity ° 8.41±1.02a 12.13 5.55±1.62b 29.19
Moisture % 45.69±1.93 4.22 46.90±2.27 4.84
Protein % 5.25±0.91 17.33 5.21±0.55 10.56
Glucose mg/kg-dw 1.17±0.32 27.35 1.57±0.77 49.04
Fat mg/kg 0.91±0.31 34.06 0.96±0.20 20.83
Salt mg/kg 2.25±0.37 16.44 2.58±0.51 19.77
Cu mg/kg 1.86±0.52a 27.96 1.44±0.17a 11.80
Mn mg/kg 18.49±9.32a 50.40 11.24±3.49b 31.05
Fe mg/kg 25.15±11.53 45.84 17.58±3.06 17.41
Zn mg/kg 17.60±8.24a 46.82 10.67±1.33b 12.46
Mg mg/kg 451.42±187.83 41.61 330.77±54.31 16.42
Ca mg/kg 403.59±246.93 61.18 321.40±81.30 25.29
−
n – samples number; × ± SD - average value; SD – standard deviation; a,b – statistically significant difference at the level p<0.05; RSD - relative standard deviation.

Health quality yeast fermentation had an important role in con-

trolling the formation of acrylamide at low lev-
Maillard reactions are the most important els of asparagine, such as those generally pres-
chemical events occurring during the manufac- ent in bread raw materials. HEDEGAARD et al.
ture of bakery products. During these reactions, (2008) found that rosemary was suppressing
healthy and potentially-harmful compounds are acrylamide formation when added to dough pri-
produced. Among the harmful ones, acrylamide or to baking. In an experiment by MUSTAFA et al.
has received a great attention in recent years. (2009), it was found that the yield of alkali-ex-
It is classified as a ‘probable human carcino- tractable acrylamide in rye crisp bread was on
gen’ and defined as a compound with the poten- average 37% higher than the yield of water-ex-
tial to cause a spectrum of toxic effects. Acryla- tractable acrylamide. The storage of samples for
mide was observed to form mainly from aspar- 20 months led to a decrease in the acrylamide
agine and reducing sugar trough Maillard reac- content but did not change the correlation be-
tions. Several factors, such as initial concentra- tween the acrylamide yield from neutral and al-
tion of the precursors, their ratio, heating tem- kaline extraction. According to research con-
perature, time of processing, pH, and water ac- ducted by CAPUANO et al. (2009), after 30 min
tivity of a product, have been reported to influ- of toasting wheat, whole-wheat and rye bread
ence acrylamide content in heated food (CLAUS samples at 180°C, the final concentration of
et al., 2008; HEDEGAARD et al., 2008; CAPUANO acrylamide was 262.3, 291.0 and 301.0 µg/kg,
et al., 2009; GÖKMEN et al., 2009; BARTKIENE respectively. The higher concentration of acryl-
et al., 2012; SERPEN et al., 2012). amide in rye bread is due to a higher free aspar-
Statistically significant differences in acryl- agine content in rye flour, which is one of the
amide content (Table 1) were recorded between factors influencing acrylamide amount in bakery
bread from small-scale and large-scale produc- products. A similar correlation between free as-
tion. A significantly lower content (P ≤ 0.05) was paragine level of flour and acrylamide concentra-
found in samples of traditional bread (119.62 ± tion in rye bread slices was reported by GRANDY
37.11 µg/kg of bread) as compared to the con- et al. (2009). MUSTAFA et al. (2008) demonstrat-
ventional samples (163.35 ± 26.22 µg/kg of ed that the concentration of acrylamide was af-
bread). fected by asparagine content. In bread produced
SVENSSON et al. (2003) showed that the con- from flour with a lower (0.44 g/500 g flour) con-
tent of acrylamide in soft wheat bread was in tent of asparagine, the content of acrylamide was
the range of < 30-160 µg/kg of fresh bread. Ac- less than 50 µg/kg, whilst a higher concentra-
cording to MUSTAFA et al. (2009), the main pre- tion of asparagine (3.08 g/500 g flour) raised
cursors of acrylamide in cereal products are as- acrylamide content to 600 µg/kg product. Ac-
paragine and reducing sugars, with asparagine cording to the International Agency on Research
being the limiting precursor. It was shown that on Cancer (1994), acrylamide was classified as

130  Ital. J. Food Sci., vol. 25 - 2013

a potential factor likely to accelerate cancer de- RSD = 45.84, for magnesium SD = 187.83, RSD
velopment in man. And although bread does not = 41.61, for calcium SD = 246.93, RSD = 61.18).
pose great risk to man, owing to a relatively low Some data suggest that sourdough bread is a bet-
content of this compound compared to French ter source of magnesium, iron and zinc compared
fries, chips or crackers, it is consumed every day to the breads produced with other methods (LOPEZ
and therefore measures should be undertaken et al., 2003; CHAOUI et al., 2006). A study by DEMI-
to reduce acrylamide content of bread. ROZU et al. (2003) showed that the mean values of
The average content of chlorinated organic iron, copper, zinc, lead and cadmium levels were
pesticides (Σ DDT, γ HCH) and polychlorinated 19.2 ± 8.1 mg/kg-dry weight (dw), 2.1 ± 1.0 mg/
biphenyls (PCBs) was low in both traditional and kg-dw, 10.0 ± 3.0 mg/kg-dw, 86.8±176.0 mg/kg-
conventional bread. No differences were report- dw, and 12.2 ± 6.1 mg/kg-dw, respectively.
ed in this respect between samples of traditional
and conventional bread (P ≤ 0.05). No residuesof Acidity, salt and moisture content
N-methylcarbamates or ochratoxin A were found
in the analyzed samples. These results seem to The samples’ place of origin (small or large
suggest that, in the analyzed region, N-methyl- companies) had no impact on values determined
carbamates and OTA contamination is well un- for concentration of moisture and salt. Acidity of
der control and unlikely to represent a threat to rye bread in samples of traditional bread (table
consumer health. It has been found that Ochra- 1) was significantly higher (8.41° ± 1.02º of acid-
toxin A (OTA) has a variety of potentially deadly ity), compared to the samples of mass-produced
toxic effects. Bread samples collected from com- bread (5.55° ± 1.62º of acidity). The results sug-
mercial outlets across Portugal were tested for gest that the production process of rye bread
OTA and all samples were found to be compli- made on a large scale does not include the fer-
ant with the European Commission. OTA con- mentation phase or that the phase was shortened
tent reached a maximum of 0.49 ng/g in Algarve in order to, for example, reduce production costs.
and 0.43 ng/g in Bragança, and was thus below
the 3 ng/g maximum limit established by Euro-
pean legislation for bread (BENTO et al., 2009). CONCLUSIONS

Nutritional value In general, the health quality and nutrition-

al value of rye bread produced by micro-com-
The average protein content in the bread un- panies localized in the province of Warmia and
der study was 5.24 ± 0.82% – 5.25 ± 0.91% in Mazury (Poland), compared to bread of the same
samples of traditional bread and 5.21±0.55% in kind manufactured in large bakeries, varied only
those of conventional bread. A study by MICHAL- with respect to some determinants. The only ex-
SKA et al. (2008) showed that both carbohydrate ceptions were the content of acrylamide, cop-
and protein concentrations were affected by mill- per, manganese and zinc. The reported levels
ing and baking processes. Protein content de- of acrylamide were relatively low to pose health
creased from 9.34 ± 0.04 to 7.85 ± 0.02 g/100 risk. The acidity of bread produced by large bak-
g of flour sample during milling, thus lowering eries suggests that it does not have the features
its nutritional quality. These results may be ex- which are typical of bread made from sourdough,
plained by a loss of grain material containing which may imply that the producers who claim
protein during milling. Baking did not affect the their bread is made with traditional methods are
protein concentration as much as milling did. misleading consumers.
The obtained data did not correspond with re-
sults reported by KOPEĆ et al. (2011). Those au-
thors showed that baking technology impacted ACKNOWLEDGEMENTS
protein quality. The differences in protein qual-
ity were due to the baking technology applied.
This study was financially supported by the Polish Minis-
The content of glucose (1.17±0.32 mg/kg-dw) try of Science and Higher Education from sources for sci-
and fat (0.91±0.31 mg/kg) in traditional bread ence in the years 2008-2011 under Research Project No. N
was similar to that in bread produced by large N312 261035.
bakeries. LAPPI et al. (2010) reported lower pro-
portions of fibre (3.3) and fat (1.5) in a white
wheat bread portion compared with wholemeal REFERENCES
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Paper received June 18, 2012 Accepted October 30, 2012

132  Ital. J. Food Sci., vol. 25 - 2013

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