Advances in Engineering Research, volume 151

International Conference on Smart Solutions for Agriculture (Agro-SMART 2018)

Stability of Flour Mixture under Various Storage

Conditions with Addition of Fenugreek Seed Powder

Lyudmila V. Anisimova Osama I. A. Soltan

Department of Grain Storage and Processing Technology PhD Student, Altai State Technical University
Polzunov Altai State Technical University Department of Food Science
Barnaul, Russia Minia University
[email protected] El-Minia, Egypt
[email protected]

Abstract—This paper investigates storage stability (165-day and have antidiabetic, hypocholesterolemic, and galactagogue
period) of whole oat flour obtained by hydrothermal treatment effects when added to some food like bread. Fenugreek is also
and stored at 40 degrees Celsius in cloth bags and flour mixture considered to be a valuable source of food proteins (70%),
containing 66.7 percent of whole oat flour obtained by good quality protein (20–25%), lysine (5–6%), and minerals,
hydrothermal treatment, 13.3 percent of fenugreek seed powder, including iron, calcium, and beta-carotene [3]. Fenugreek seed
6 percent of dry gluten, and 14 percent of wheat flour stored at is a good source of dietary fiber as well. This fiber functional
20 and 40 degrees Celsius in various types of bags (namely, cloth ingredient could be included in flour mixture production.
and plastic). Measurement of moisture content, acidity, acidity of
flour by water-alcohol extraction, an acid number of fat as well Fenugreek seed powder is aromatic and contains
as microbiological load was carried out during the storage antibacterial and bioactive components [4]. Thus, addition of
period. The results revealed that at all storage conditions with the fenugreek seed powder extends shelf life of products.
increase of storage time flour acidity, acidity of flour by water-
alcohol extraction, and an acid number of fat increased, whereas To increase the consumption of staple food made from
microbiological load decreased. Changes in moisture content of oats, we should expand their range [5, 6, 7]. Still, in some
stored samples are affected by various temperatures and in cases, the use of oat is limited due to its tendency to rancidity
different types of bags. However, flour mixture stored at 20 that causes a bitter taste and rancid smell in its by-products
degrees Celsius remained shelf stable over the time storage [8]. The development of rancidity is particularly intense in the
period. Whole oat flour stored at 40 degrees Celsius in cloth bags production of unprocessed "raw" oats. Its prevention is usually
showed larger increase in the acidity, acidity of flour by water- achieved through the application of heat combined with high
alcohol extraction, and an acid number of fat. Flour mixture humidity required for the inactivation of endogenous lipolytic
containing fenugreek seed powder and stored in plastic bags saw enzymes in oats [9, 10].
lower increase in acidity and an acid number of fat than the flour
mixture in cloth bags and whole oat flour at 40 degrees Celsius. During the storage period, there occurred various chemical
processes, including the accumulation of free fatty acids and
Keywords—whole oat flour; storage stability; fenugreek seed further peroxides, in cereal grain products. These processes
powder; acidity; moisture content; acid number of fat develop at a tremendous rate and lead to rapid deterioration in
product quality. An acid number of fat can be an indicator of
I. INTRODUCTION biochemical changes taking place during the storage of cereal
Oats (Avena sativa) is a grain crop grown around the products. Fat acidity is defined as the number of milligrams of
world. Products processed from oats not only are the source of potassium hydroxide required to neutralize the free fatty acids
essential nutrients, such as starch, protein, fats, vitamins, from 100 g of cereal products and calculated to a moisture-
minerals, etc., but also play an important role in the prevention free basis. Under the Codex Alimentarius, the limit of an acid
of chronic diseases of diabetes, coronary heart disease, and number of fat (50 mg KOH/100 g ) was applied only to white
cancer. Healthy properties of oats are attributed to the flours of extraction rates up to 75%.
presence of a number of biologically active compounds found Flour from cereal crops is generally regarded as a
in different parts of the grain. One of these compounds is microbiologically safe product owing to its low humidity [11].
soluble fiber beta-glucans contained in cell walls of the However, microbiological control of flour quality during its
endosperm [1]. storage is necessary as it can trap microorganisms that cause
Fenugreek (Trigonella foenum-graecum) was originally food spoilage.
cultivated in South East Europe and West Asia, but now it is Therefore, the aim of this study is to investigate the effects
mainly grown in Egypt and India [2]. The fenugreek seeds are of storage time, storage temperature, package type, and
primarily used as spice for flavoring Egyptian food products

Copyright © 2018, the Authors. Published by Atlantis Press.

This is an open access article under the CC BY-NC license (http://creativecommons.org/licenses/by-nc/4.0/). 29
 Advances in Engineering Research, volume 151

addition of fenugreek seed powder on the chemical changes D. Microbiological Load

and microbiological load of whole oat flour and flour mixture
based on whole oat flour. Besides, we compared analyses of The microbiological analysis (total bacteria) of oat flour
acidity, moisture content, and fat acidity as part of our study. was carried out immediately after production and during its
storage. Each sample (1 g) was serially diluted by dissolving
II. RESEARCH METHODS in 9 mL of distilled water. One milliliter from each diluent
was plated in Petri dish and plated out on nutritional agar
A. Materials plates for bacterial and fungal count. The plates were
Oat Samples. The oat samples (Avena sativa L.) were incubated at 37°C for 48 h for bacterial growth and 7 days for
grown in 2017 in Altai region. The samples were carefully fungal growth [16].
cleaned, treated with hydrothermal processing, and then milled
by a laboratory mill 3100 (Finland) into fine powder. At the
Department of Grain Storage and Processing Technology, Raw material (Oat grain)
Altai State Technical University the technology of the
production of flour from oat is based on the use of
hydrothermal treatment. In the experiment, the hydrothermal
treatment of the oats was carried out as follows: first, the grain Moistening under vacuum
was moistened in a laboratory unit by adding a calculated
amount of water under vacuum. Then, it was put into
hermetically sealed containers for a predetermined time. Resting
Further, the grain was dried to predetermined moisture in a
laboratory drier in a stream of heated air.
Wheat Flour and Fenugreek. Wheat flour samples were Drying
obtained from the local market in the city of Barnaul, Russia,
whereas the fenugreek was obtained from the local market in
Dehulling
Egypt. The fenugreek samples were carefully cleaned and then
milled by a laboratory mill into fine powder.
B. Storage Studies of Oat Flour Milling
Flour mixtures containing 66.7% of whole oat flour
obtained by hydrothermal treatment,13.3% fenugreek seed Whole oat flour
powder, 6% dry gluten, and 14% wheat flour (Fig. 1) were
properly weighed (45 g each), packaged in cloth bags, closed,
and stored at 20±0.5°C and relative humidity 65±0.1%. 6% 66.7 % 13.3% 14 %
Additionally, flour mixtures to be stored at 40±0.5°C were Dry Whole Fenugreek Wheat
divided into two portions: the first one was packaged in cloth
gluten oat flour seed powder flour
bags, while the second one was packaged in zip lock plastic
bags. Whole oat flour in cloth bags was then closed to
determine the storage stability. The temperatures and relative
humidity inside the storage box were measured with a thermo-
hygrometer before sample collection. Storage at 40⁰ C Storage at 20⁰ and 40⁰ C
Moisture content, acidity, acidity of flour by water-alcohol
extraction, and fat acid number of the samples were identified Fig. 1. Flow chart of whole oat flour obtained and mixture preparation
every 2 weeks, while the microbiological counts were done
every 4 weeks of storage. III. RESULTS AND DISCUSSION
C. Chemical Analysis A. Moisture Content
 Moisture content was determined by using an air Fig. 2 shows moisture content of whole oat flour stored at
oven at 130°C / 40 min according to GOST 9404- 40°C in cloth bags. From these results we can see that
moisture content of whole oat flour decreased during the first
88 [12]. storage period within two months. This decrease of moisture
 Acidity was determined by the calibration method content may be attributed to the equilibration process of the
according to GOST 27493-87 [13]. stored flour with the surroundings [17]. After that, there was a
 Acidity of flour by water-alcohol extraction was slight increase in the moisture content that later stabilized due
to the equilibrium between the moisture content of flour and
determined by the calibration method [14]. the relative humidity of storage environment.
 Acid number of fat was determined according to
Fig. 2 also demonstrates moisture content for flour
GOST 31700-2012 [15]. mixtures. The data indicate that moisture in flour mixture
rapidly increased during the first month, after that the chart

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shows a slight decrease that is followed by the increase during During the storage, the acidity of all samples of whole
the second month that further on stabilizes until the end of the flour and flour mixture increase may be attributed to the
storage period. The increase in the moisture content of flour accumulation of linoleic and linolenic acids [19]. It is further
mixture can be attributed to the addition of fenugreek seed to mention that during the whole storage period the acidity and
powder, dry wheat gluten, wheat flour and its hydrophilic acidity by water-alcohol extraction of the whole oat flour
compounds effect on water absorption of flour mixture [18]. stored at 40°C in cloth bags showed higher titratable acidity
Higher moisture content was indicated in flour mixture stored and acidity by water-alcohol extraction rate as compared to the
in cloth bags at 20°C. However, the type of bags did not acidity of flour mixture.
produce a significant effect on moisture content of flour
mixture stored at 40 °C. The study found out that the addition of fenugreek seed
powder in flour mixture results in a slight increase in the
B. Acidity and Acidity by Water-Alcohol Extraction samples acidity and acidity by water-alcohol extraction under
The acidity and acidity by water-alcohol extraction of various storage conditions during the storage period. This can
grain and flour characterize their freshness. When grain and be explained by the ability of fenugreek seed powder to reduce
products of its processing are stored, their acidity, as a rule, the degradation of the compounds of flour mixture, such as
increases. protein, and thus to reduce its acidity – that corresponds to the
results found by [20]. Obviously, the lower level of acidity
and acidity by water-alcohol extraction of flour mixture stored
at 20°C may be explained by the slight effect of temperature
on degradation of chemical structure of flour mixture and the
partial inactivation of enzymes.
The study also examined flour mixture stored at 40°C in
two types of bags (cloth and plastic). The results showed that
acidity and acidity by water-alcohol extraction rises at a lower
rate in the samples stored in the plastic bags compared to the
samples in the cloth bags. The reason is that the plastic bags
reduce the exposure of oxygen to mixture samples, thus
reducing the oxidation and degradation of flour mixture
components that results in the reduction of the acidity and
acidity by water-alcohol extraction increased during the
storage period.
C. Acid Number of Fat
Acid number of fat is another indicator of the degree of
freshness of grain and products of its processing; its increase
eventually leads to the development of rancidity and food
spoilage [21].
Fig. 2. Effect of storage conditions on moisture content Fig. 3 shows that the rate of change in the acid number of
fat in the whole oat flour stored at 40°C can be described as
three stages. The first stage (up to 30 days) was an induction
The increase in the acidity and acidity by water-alcohol period when the rates changed slightly; the second stage was
extraction of flour is caused by the accumulation of products the development process (from 30 to 90 days) characterized by
of hydrolytic breakdown of fats, phosphorus compounds, and the significant change in biochemical parameters, while the
carbohydrates. third stage can be considered as the stage of decay (90-165
Table 1 shows that in zero time the titratable acidity and days). Increase in the acid number of fat during the storage
acidity by water-alcohol extraction of the whole oat flour might be attributed to the activity of enzymes, such as lipases,
obtained from the oat grain after hydrothermal treatment rate lipoxygenase and acylhydrolases in whole oat flour [22]. The
was significantly less than the acidity of flour mixture after storage stability of whole oat flour and its acid number of fat
addition of fenugreek seed powder and dry gluten. was also affected by the storage temperature (40°C) that led to
the degradation of lipid.
The lower rate of the whole oat flour acidity and acidity by
water-alcohol extraction as compared to the flour mixture is However, the stages were different in case of flour mixture
accounted for the various chemical compositions of these stored at 20°C. Two stages can be identified here: the first
products. At the same time, the effect of heat and moisture in stage (up to 105 days) was an induction period when the rates
the hydrothermal treatment of grain leads to partial changed slightly and acid number of fat increased from 17.4
denaturation of proteins and a decrease in the activity of mg of KOH/ 1 g of fat to 40 mg after 105 days of storage
enzymes that also contributes to the decrease in the acidity of respectively; the second stage was the development process
oat flour.

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(from 120 to 165 days) when an acid number of fat increased degradation and a decrease in an acid number of fat under
from 53 mg of KOH/ 1g of fat to 73.5 mg after 165 days of these storage conditions.
storage respectively. This indicates a decrease in the rate of fat

TABLE 1. EFFECT OF STORAGE CONDITIONS ON ACIDITY AND ACIDITY BY WATER-ALCOHOL EXTRACTION

Storage samples
Storage Whole oat flour cloth Flour mixtures cloth bags Flour mixtures plastic Flour mixtures cloth bags
period bags 40 °C 20°C bags 40°C 40°C
(days) Acidity by Acidity by Acidity by Acidity by
Acidity a water-alcohol Acidity a water-alcohol Acidity a water-alcohol Acidity a water-alcohol
extraction b extraction b extraction b extraction b
Zero time 3.1 2.0 3.9 2.5 3.9 2.5 3.9 2.5
15 5.5 5.0 4.2 3.5 4.5 4.0 4.9 4.5
30 5.7 5.5 4.6 4.0 5.1 4.5 5.6 5.0
45 6.1 5.5 5.1 4.5 5.6 5.0 5.9 5.5
60 6.2 5.9 5.2 4.9 5.7 5.5 6.0 5.5
75 6.3 6.0 5.4 5.0 5.8 5.7 6.1 5.9
90 6.5 6.0 5.7 5.4 6.0 6.0 6.3 6.4
105 6.8 6.5 6.2 6.1 6.4 6.6 6.4 6.8
120 7.5 7.5 6.4 6.3 6.7 7.0 6.9 7.1
135 8.0 7.7 6.5 6.2 7.1 7.1 7.5 7.4
150 8.9 8.2 7.0 7.0 7.3 7.2 7.7 7.5
165 9.8 10.5 8.1 7.5 8.5 8.2 8.6 9.2

a. Degree of acidity.
b. Acidity degree by water-alcohol extraction.

Furthermore, the results showed that the addition of same temperature (40°C). It stems from the fact that the
fenugreek seed powder to flour mixture significantly decreases fenugreek seed powder contains flavonoids, such as isovitexin
the rate of increase of an acid number of fat when using both and vitexin that have the antioxidant effect [23]. However, the
types of bags (cloth and plastic). This bioactive effect of antioxidant effect lasted only until the 135th day of storage.
fenugreek seed powder causes longer preservation of flour Then acid number of fat increased due to the deterioration of
mixture without oxidation and degradation of the lipids flavonoid compounds in the fenugreek seed stored at high
compounds as compared to the whole oat flour stored at the temperature (40°C) for a long period of time.

Fig. 3. Effect of storage conditions on acid number of fat

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The results showed that on the one hand, the type of Absence of fungi growth in both whole oat flour and flour
storage bags (cloth or plastic) did not produce a significant mixture arises from the thermal treatment and vacuum
effect on the acid number of fats in flour mixtures stored at treatment used to obtain whole flour by hydrothermal
40°C for the antioxidant effect of fenugreek seed powder. We treatment and killing the fungi as well as the addition of
did not observe significant high oxidation effect on flour fenugreek seed powder in flour mixture with its inhibitory
mixture stored in the cloth bags until the 75th day of effect on fungi growth.
storage. However, the increase in the storage period at high
The results showed that in case of whole flour mixtures
temperature led to the degradation of flavonoid
stored at 40°C in cloth bags the bacterial count increased in the
compounds (antioxidants).
first period of storage (until 30 days). Still, in the later stages
Therefore, the type of bags had a strong effect on acid the bacterial count minimized because the acidity of whole oat
number of fat. It was recorded that the rate of increase in fat flour increased and inhibited bacterial growth [24].
acidity was higher in cloth bags as compared to samples stored
in plastic bags. The obtained result may be attributed to the The study found out that the bacterial count in all flour
ability of plastic bags to isolate the air streams and reduce the mixture samples with fenugreek seed powder decreased since
lipid oxidation. fenugreek shows antifungal and antibacterial activity [25].
However, the bacterial count slightly decreased in flour
D. Microbiological Load
mixture stored at 20°C as compared to flour mixtures stored at
Fig. 4 illustrates the total bacterial count in whole oat flour 40°C. These results correspond to the results of acidity
stored in cloth bags and in flour mixtures with fenugreek seed measurement when the acidity level of samples stored at 40°C
powder stored at 20 and 40°C in cloth and plastic bags. increased more than that of the mixture stored at 20°C. On the
According to the results, the initial storage period showed no other hand, the type of bags showed no significant difference
significant difference between the bacterial count in whole oat in bacterial growth rate.
flour obtained by hydrothermal treatment and flour mixture. In
addition, fungi growth was not recorded in this study.

Fig. 4. Effect of storage conditions on microbiological load

IV. CONCLUSION flour at the same storage temperature because the oxidation
activity causing rancidity was inhibited by antioxidant
This study demonstrates factors that affect shelf stability of compounds in fenugreek seed powder. The study found out that
whole oat flour and flour mixture, namely storage temperature, the bacterial count in all flour mixture samples containing
bags type, fenugreek seed powder addition, and its importance fenugreek seed powder decreased since fenugreek showed
in extending the shelf life of flour mixture. The stability of antifungal and antibacterial activity. According to the research,
flour mixture significantly increased with the addition of fenugreek seed powder might be of critical importance in
fenugreek seed powder at 20 and 40°C. The increase in acid extending the storage period of flour mixture.
number of fat was lower in flour mixture than in the whole oat

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