Ecological Engineering 84 (2015) 229–232

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Ecological Engineering
journal homepage: www.elsevier.com/locate/ecoleng

Short communication

Extracts of fava bean (Vicia faba L.) seeds as natural coagulants

Dragana V. Kukić a,∗ , Marina B. Šćiban a , Jelena M. Prodanović a , Aleksandra N. Tepić a ,
Mirjana A. Vasić b
a
University of Novi Sad, Faculty of Technology, Blvd. Cara Lazara 1, 21000 Novi Sad, Serbia
b
Institute of Field and Vegetable Crops, Maksima Gorkog 30, 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: In this study, applicability of extracts from fava bean seeds (Vicia faba L.), as natural coagulants, was inves-
Received 16 October 2014 tigated. Coagulants were obtained by extraction, with distilled water or NaCl solutions, from grinded seeds
Received in revised form 7 July 2015 and were applied as coagulants, in various doses, in synthetic water having different initial turbidities
Accepted 4 September 2015
and pH values. Presence of NaCl in water showed that increasing the ionic strength results in greater
amount of extracted compounds, but it does not affect coagulation activity. Appling different doses of
Keywords:
coagulant in water showed that the best coagulation activity is achieved by adding dose of 0.125 ml/l.
Natural coagulant
The conducted experiments confirm positive coagulation properties of these extracts, therefore it can be
Fava bean
Water
concluded that they have great potential as natural coagulants.
Turbidity © 2015 Elsevier B.V. All rights reserved.
Clarification

1. Introduction the aim of this study was to investigate fava bean seeds and the
applicability of their extracts as natural coagulants.
Removal of water turbidity and color, in form of suspended and
colloidal material, is generally achieved by coagulation and floc-
culation processes (Teh et al., 2014). Some of the most commonly
used coagulants are aluminum salts, but they have serious disad- 2. Materials and methods
vantages, such as, production of harmful sludge and indications that
their residues in treated water can be harmful for human health 2.1. Preparation of seed extracts and synthetic turbid water
(Jodi et al., 2012; Bondy, 2010; Betatache et al., 2014). Lately, nat-
ural coagulants are widely investigated because of their efficiency, Natural coagulants were extracted from seeds of fava bean strain
abundant source, low price and biodegradability (Antov et al., 2012; named Talijan, cultivated in the Institute of Field and Vegetable
Šćiban et al., 2009a; Betatache et al., 2014). Many materials of plant Crops, Novi Sad. The whole seeds were grinded to fine powder,
origin were studied as a source of natural coagulants (Sanghi et al., using a laboratory mill, and sieved through 0.4 mm sieve. The frac-
2002; Šćiban et al., 2005; Ghebremichael et al., 2009; Teh et al., tion, with particle size less than 0.4 mm, was used in experiments.
2014; Fedala et al., 2015). Seeds from tropical plant Moringa oleifera A solid sample of fava bean seeds was analyzed in terms of nitro-
have shown to be one of the most effective primary coagulants for gen and proteins content: 4.72% and 29.5% (expressed on the basis
water treatment (Katayon et al., 2006; Betatache et al., 2014). Faba of dry matter), respectively. 10 g/l of this fraction was suspended
bean (Vicia faba L.) is an ancient crop of high nutrition values. In in distilled water or NaCl solutions. The suspension was stirred for
Mediterranean area it has been a staple food for a long time but on 10 min on a magnetic stirrer, in order to extract active coagulants,
the Balkans it was replaced by Phaselous beans (Mihailović et al., and then filtered through filter paper Macherey-Nagel MN 651/120.
2010), which belong to the same family as fava bean – Fabaceae Obtained filtrates were stored in a refrigerator at 5 ◦ C.
(Link et al., 2008). In some rural areas it was used for water clarifi- Synthetic turbid water was prepared by adding stock kaolin sus-
cation, but this possibility was not investigated enough. Therefore, pension into tap water. At first, kaolin was grinded in a mortar and
sieved (0.4 mm sieve). A smaller fraction was taken to prepare 1%
kaolin suspension with tap water. The suspension was stirred for
∗ Corresponding author.
1 h and then it was left to rest for 24 h. Synthetic turbid water was
E-mail addresses: [email protected] (D.V. Kukić), [email protected]
prepared just before performing the coagulation test, by adding
(M.B. Šćiban), [email protected] (J.M. Prodanović), [email protected] (A.N. Tepić), this 1% kaolin suspension in 1 L of tap water, in order to obtain the
[email protected] (M.A. Vasić). water with desirable value of initial turbidity. The initial pH in the

http://dx.doi.org/10.1016/j.ecoleng.2015.09.008
0925-8574/© 2015 Elsevier B.V. All rights reserved.
230 D.V. Kukić et al. / Ecological Engineering 84 (2015) 229–232

synthetic turbid water was adjusted to a desired value with 1 M established what compound is an active component, chemical com-
NaOH or 1 M HCl. position of extracts does not give enough information about their
efficiency as natural coagulants. It is necessary to conduct coagula-
2.2. Coagulation tests tion tests. Coagulation tests for fava bean extracts were performed
in synthetic turbid water at pH 7, initial turbidity of 45 NTU and
Coagulation activity of all extracts was determined by the jar test coagulant dose of 0.5 ml/l.
using synthetic turbid water. Just before the test, pH value of syn- Proteins in all investigated extracts of fava bean seeds were
thetic turbid water was adjusted and the water (200 ml) was filled present in higher concentrations than phenolic components and
into the beakers (600 ml) and mixed at 200 rpm at room temper- phytic acid (Table 1). It was presumed that the proteins are
ature. Various doses of seed extracts were added into the beakers responsible for coagulation activity, but other compounds such
and mixed for 1 min. The mixing speed was then reduced to 80 rpm as phenolics and phytic acid may also contribute to the coagu-
and kept for another 30 min. After that, the system was left to sed- lation activity of investigated extracts. According to Crépon et al.
iment for 1 h. The same coagulation test was conducted with no (2010), fava bean seeds contain some constituents that may exert
coagulant (blank). Clarified samples (50 ml) were collected from anti-nutritional effects which include tannins, widely occurring
the top of the beakers and residual turbidity was measured using phenolic components in plants. Some studies indicate that tan-
turbidimeter. The coagulation activity was calculated as: nins can be a source of coagulation agents and that they can be
used in water treatment (Özacar and Şengil, 2002). Özacar and
(Tb − Ts ) × 100
Coagulation activity (%) = (1) Şengil (2002) investigated tannins obtained from valonia, as pri-
Tb
mary coagulants and coagulant aid, and the results showed that
where Tb is turbidity of the blank and Ts is sample turbidity. tannins operated much better as a coagulant aid than as a primary
coagulant. It may be that they have the same role in fava bean
2.3. Analytical methods extracts. Phytic acid is also known as anti-nutrient, but also as a
natural wastewater cleaner used on industrial scale (Vasić et al.,
In solid sample and crude extracts the following parameters 2012). Extracts of fava bean with NaCl solutions contained slightly
were determined: dry matter (APHA, 1998), total sugars (Official higher concentration of phenolics compounds and phytic acid than
Gazette of SFRJ, 1983), content of total nitrogen (protein content extracts of common bean analyzed by Kukić et al. (2011), and per-
was calculated as total nitrogen content multiplied by 6.25 (MEBAK, formed slightly higher coagulation activity. In water extracts of fava
1997)), total phenolics (Waterhouse, 2002), content of phytic acid bean, proteins were present in greater amount compared to litera-
(Haug and Lantzsch, 1983), pH (pH-meter Oakton Ion 6 with epoxy- ture data for common bean extracts (Kukić et al., 2011; Šćiban et al.,
body combination pH electrode), water turbidity (WTW Turb 550 2009b). Amounts of extracted proteins from fava bean seeds were
IR), expressed in nephelometric turbidity units (NTU), isoelectric higher about 24% compared to Phaselous beans (Kukić et al., 2011).
point, zeta potential and particle size distribution of the extracts Coagulation test of Kukić et al. (2011), with common bean extracts,
(Zetasizer Nano ZS, Malvern Instruments). were performed at optimal pH for those coagulants (pH 9), initial
turbidity of 35 NTU and coagulant dose of 0.5 ml/l. Even though ini-
3. Results and discussion tial turbidity in coagulation tests presented in this study, with water
fava bean extracts, was higher (45 NTU), these extracts showed the
3.1. Analyses of the solid sample of fava bean same coagulation activity as water extracts of common bean. Other
two extracts of fava bean showed higher coagulation activity com-
It was suggested that proteins are active components in plant pared to corresponding extracts of common bean. Water extract of
extracts, responsible for coagulation processes (Ndabigengesere soybean, investigated by Šćiban et al. (2009b), contained less pro-
et al., 1995; Ghebremichael et al., 2006). Compared to typical data of tein and had lower coagulation activity at initial turbidity of 35 NTU
chemical composition of fava bean, it can be concluded that inves- and dose of 0.5 ml/l, than fava bean extract. On the basis of obtained
tigated species had high content of proteins (29.5%), but still in results in this study, it was concluded that higher ionic strength
range characteristic for fava beans. Makkar et al. (1997) analyzed results in better extraction of all components from fava bean seeds.
12 different cultivars of fava bean and the content of proteins varied The greatest amount of proteins, total sugars, phytic acid and phe-
from 25.6% to 30.4%. Compared to other leguminous species, inves- nolic compounds were extracted with 1 M NaCl solution. The same
tigated by Šćiban et al. (2009b), solid sample of fava bean contains tendency was noticed in extraction of proteins and phenolic com-
less nitrogen than soybean (about 20%), but more than Phaselous pounds from common bean seeds, with solutions of different ionic
beans (about 24%). strength, while the content of total sugars decreased (Kukić et al.,
2011). Compared to mentioned results for common bean extracts,
3.2. Analyses of the extracts of fava bean the content of all compounds was higher in extracts of fava bean
seeds. Coagulation activities of fava bean extracts with water and
The active components from grounded solid samples can be 1 M NaCl were similar but the coagulation activity of 0.5 M NaCl
extracted from seeds by water, different salt solutions, buffer extract was lower, about 25%. Therefore, it was recommended to
solutions or organic solvents (Ndabigengesere et al., 1995; extract the active components from fava bean seeds, by distilled
Ghebremichael et al., 2006). In this paper distilled water, 0.5 M water, and so all further experiments, in this paper, were conducted
NaCl and 1 M NaCl were used in order to determine their influence with water extracts.
on the quantity of extracted components. Obtained extracts were
analyzed and content of proteins (mg/l), phytic acid (mg/l), total 3.2.1. Isoelectric point (IEP) and zeta potential of fava bean
phenolics (mg/l) and total sugars (mg/l) were determined. Each extract
analysis was carried out in triplicate and the results were averaged. Isoelectric point is the pH value of a particular molecule, at
Some authors suggest that active coagulation agents in extract which it has no electrical charge. IEP can affect the solubility
from M. oleifera seed are dimeric cationic proteins (Ndabigengesere of a protein. Proteins have minimum solubility in water or salt
et al., 1995; Fink, 1984). Sanghi et al. (2002) and Okuda et al. (2001) solutions at the pH that corresponds to their IEP and often precip-
suggested that the active component is not protein, polysaccharide itate out of solution. For this reason it is necessary to determine
nor lipid, but some other organic polyelectrolyte. Since it was not IEP of extracts and optimal pH value for further experiments.
 D.V. Kukić et al. / Ecological Engineering 84 (2015) 229–232 231

Table 1
Content of some components in fava bean and common bean extracts and their coagulation activity.

Source Fava bean Common bean

(present study) (Kukić et al., 2011)

Distilled water 0.5 NaCl 1 M NaCl Distilled water 0.5 NaCl 1 M NaCl

Proteins (mg/l) MV 2460 3423 3828 1909 1860 2034

 76 76 109 – – –
Phytic acid (mg/l) MV 20.8 34.8 35.9 14.5 15.2 11.7
 2.5 0.4 2.5 – – –
Total phenolics MV 1.93 2.06 2.72 1.11 1.16 1.20
content (mg/l)  0.37 0.29 0.49 – – –
Total sugars MV 729 578 1076 697 553 406
content (mg/l)  15 169 34 – – –
Coagulation activity (%) MV 33.0 24.8 34.8 33.0 20.3 30.7

MV – mean value.
 – standard deviation.

Fig. 1. Zeta potential in extract of fava bean at pH 7.

Consequently, in water extract of fava bean seeds the isoelectric very low coagulation activity in range from pH 5 to 8 (Šćiban et al.,
point and zeta potential (Fig. 1) were determined, as well as size 2009a). As shown in Section 3.2.1, particles of different sizes and
distribution of particles in suspension by volume. electrical charge were present in extracts and it was assumed that
Results showed that the isoelectric point of particles in sus- all of them participate in coagulation process, which can be the
pension was at pH 4.04 and that the particles with diameter of cause of poor influence of pH on clarification efficiency. Because of
39.85 nm occupied 6.9%, particles with diameter of 198.4 nm occu- that, further experiments were conducted at neutral pH.
pied 48.7% and particles with diameter of 5590 nm occupied 44.4%
of total volume. Zeta potential and size distribution of particles sug- 3.3.2. Effect of coagulant dose and initial water turbidity
gest that, in this suspension, particles of different electrical charge Synthetic turbid water of different initial turbidities (20, 45 and
and dimensions are present. 90 NTU) was prepared as described in Section 2.2. All tests were
conducted at pH 7. Various doses of extract were applied: 0.05,
3.3. Coagulation activity of fava bean extract 0.125, 0.25, 0.5, 1 and 1.5 ml/l, in synthetic water of all three tur-
bidities. Coagulation activities are shown at Fig. 3.
3.3.1. Effect of pH value of synthetic turbid water Better coagulation activity was achieved in water with higher
Coagulation process is affected by many parameters, such as turbidity. In synthetic turbid water with lowest initial turbidity
composition and dose of coagulant, initial water turbidity, pH, tem- (20 NTU), the best coagulation activity was achieved with extract
perature, composition of water, etc. If the pH value of surrounding dose of 0.25 ml/l (38%). At higher doses applied, coagulation activ-
environment is distant form isoelectric point, proteins become ities decreases. At dose lower than 0.25 ml/l, lower coagulation
more positively or negatively charged and more stable. It can be activity was also observed. In water with initial turbidity of 45 NTU,
expected that pH value of synthetic turbid water affects stability
and coagulation activity of proteins extracted from fava bean seeds.
Therefore, coagulation activity was investigated at different pH val-
ues (3–10) and at initial turbidity of 45 NTU, with the same dose of
the extract (0.5 ml/l) by conducting coagulation tests.
The results are shown in Fig. 2.
pH of synthetic turbid water had no significant influence on
coagulation activity in a wide range of pH values. Since the IEP is
at pH 4.04, it would be expected that the extract has no coagula-
tion activity. Its content is very complex and different compounds
can be coagulation agents at different pH values, which can cause
this kind of behavior. In similar experiments, with extracts of horse
chestnut and acorn, coagulation activity was the best at pH 3–4
and 8–10 and lower at neutral pH. The most appropriate pH for
these extracts to perform coagulation, was pH 10 and they showed Fig. 2. Effect of pH on coagulation activity of fava bean extract.
232 D.V. Kukić et al. / Ecological Engineering 84 (2015) 229–232

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