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Ecological Engineering
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Article history: Efficiency of natural coagulant obtained by ultrafiltration of common bean (Phaseolus vulgaris) seed extract
Received 9 March 2012 in water turbidity removal was experimentally evaluated. Prepared ultrafiltration fractions have shown
Received in revised form 6 July 2012 differences in behavior regarding pH values and dosages at which they expressed their highest efficiency
Accepted 10 August 2012
in turbidity removal. The highest obtained coagulation activities were at pH 9.5 with crude extract and
Available online 28 September 2012
fraction containing molecules with molecular weight less than 10 kDa, 50.6% and 49.1%, respectively. The
organic load that remained in water after treatment was reduced when natural coagulant processed by
Keywords:
ultrafiltration instead of crude extract was applied. Moreover, fraction with molecules between 10 kDa and
Natural coagulant
Ultrafiltration
30 kDa, decreased chemical oxygen demand by 9% relative to blank at conditions this fraction expressed
Coagulation activity its highest coagulation activity.
Organic load © 2012 Elsevier B.V. All rights reserved.
Common bean seed
0925-8574/$ – see front matter © 2012 Elsevier B.V. All rights reserved.
http://dx.doi.org/10.1016/j.ecoleng.2012.08.015
M.G. Antov et al. / Ecological Engineering 49 (2012) 48–52 49
separation technique was applied instead of isolation and purifi- stirred at 200 rpm for 1 min. The mixing speed was then reduced
cation multiple steps procedure to examine its suitability for the to 60 rpm and was kept for 30 min. Then, the suspensions were
processing of natural coagulant. Coagulation efficiencies of pre- left to allow sedimentation. After 1 h of sedimentation, upper clar-
pared fractions were measured at different pH values and dosages, ified liquid was collected and residual turbidity was measured. The
and have been compared to the one of crude extract. In addition, residual turbidity of sample was RTS . The same coagulation test was
suitability of prepared natural coagulants was tested in respect of performed with no coagulant as the blank. The residual turbidity
organic load that remained in water after treatment. in the blank was RTB . Coagulation activity was calculated as:
RTB − RTS
2. Materials and methods Coagulation activity (%) = · 100 (1)
RTB
2.1. Extraction of active components from common bean seed
2.5. Analytical methods
The locally obtained common bean (P. vulgaris) dry seed was
Protein concentration was measured according to Bradford
ground to a fine powder by using a laboratory mill and sieved
(1976) with bovine serum albumin as standard. Turbidity was mea-
through 0.4 mm sieve. The fraction with particle size less than
sured using a turbidimeter (WTW TURB 550/550 IR) and it was
0.4 mm was used in experiments. Ten grams of seed powder
expressed in nephelometric turbidity units (NTU). Chemical oxygen
was suspended in 1 L of NaCl water solution (0.5 mol/L). The
demand was determined according to Standard Methods (APHA-
suspension was stirred using a magnetic stirrer for 10 min to
AWWA-WEF, 1998).
accomplish extraction and then filtered through a rugged filter
All experiments were run in duplicate (the accuracy is consid-
paper (Macherey-Nagel, MN 651/120) to obtain filtrate – crude
ered to be ±5%) and the mean value is presented herein.
extract of active components.
60 60
Crude extract st
1 fraction
50 50
pH 9.0
pH 9.5
Coagulation activity (%)
30 30
20 20
10 10
0 0
0.0 0.5 1.0 1.5 2.0 2.5 0.0 0.5 1.0 1.5 2.0 2.5
60 60
nd rd
2 fraction 3 fraction
50 50
Coagulation activity (%)
Coagulation activity (%)
40 40
30 30
20 20
10 10
0 0
0.0 0.5 1.0 1.5 2.0 2.5 0.0 0.5 1.0 1.5 2.0 2.5
Fig. 1. Coagulation activities of crude extract from common bean seed and fractions prepared by ultrafiltration in relation to coagulant dosages and at different pH values.
et al., 2005). Residual turbidity of the blank varied slightly within resulting in coagulation activity just below 50%, the 1st fraction
investigated pH range and its mean value was 23.4 ± 0.6 NTU. has shown also at pH 9.5 but at double dosage in comparison to
Crude extract from common bean seed as well all ultrafiltra- crude extract. In addition, at pH 10 this fraction containing com-
tion fractions have shown differences in behavior when they were pounds having molecular weight below 10,000 Da has also showed
evaluated for turbidity removal regarding pH and dosages at which high coagulation activity in comparison to other results, 47.6% with
they expressed the highest obtained coagulation activities as well relatively low dosage 0.5 mL/L.
as the values of this highest activities. These differences might be In comparison to crude extract and other two ultrafiltration
explained by differences in kind and characteristics of biomolecules fraction, the highest coagulation activity of the 2nd fraction was
as well as by their content in fraction and crude extract. Proteins the lowest one, 45.3%. It was achieved at the highest investigated
as well as other extracted molecules separated by ultrafiltration pH value, pH 10, which appeared to be the most appropri-
differ not only in molecular weight but e.g. in pI which may addi- ate for molecules contained in this fraction to act as turbidity
tionally affect coagulation as consequence of their charge at certain removing agents. However, at the same conditions, this frac-
pH. However, it can be noticed that the higher efficiencies in tur- tion exhibited the best performance regarding COD removal (see
bidity removal generally were obtained at higher investigated pH Table 1). As for the 3rd fraction containing biomolecules above
values and that pH 9 was the least appropriate for common bean 30 kDa and having the highest protein concentration, maximum
seed molecules to express their coagulation action. of coagulation activity was recorded at pH 9.5 with relatively
Crude extract has attained its highest coagulation activity, small volume of fraction added to turbid water, 0.25 mL/L. In
slightly above 50%, at dosage 1 mL/L and pH 9.5. In comparison comparison to this result, the highest coagulation activities this
to this result, highest obtained coagulation activities crude extract fraction has shown at pH 9 and pH 10 were about 2.4 and 1.4
has reached at pH 9 and 10 were about 2.4 and 1.5 times lower, times lower, respectively, which were achieved at higher dosage,
respectively. The highest obtained efficiency in turbidity removal 1 mL/L.
M.G. Antov et al. / Ecological Engineering 49 (2012) 48–52 51
Natural coagulant Crude extract 1st fraction 2nd fraction 3rd fraction 4. Conclusion
Dosage (mL/L) 1.0 2.0 1.0 0.25
pH 9.5 9.5 10.0 9.5 Evaluation of fractions obtained by ultrafiltration of crude
Coagulation activity (%) 50.6 49.1 45.3 47.7 extract of common bean seed for their suitability for turbid-
COD (mg O2 /L) 3.74 3.33 1.84 2.32
ity removal revealed that they were efficient in it although
their coagulation activities were slightly lower that of crude
Evaluation of crude extract of common bean and ultrafiltration extract. Obtained fractions brought reduced organic load into
fraction for turbidity removal revealed that the most efficient was the water after the treatment in comparison to crude extract or
the crude extract. Although lower than that of crude extract, the even diminished it in comparison to blank. According to these
highest coagulation activities of all fractions were close to this high- results ultrafiltration appeared to be promising technique for
est obtained result. Efficiency of crude extract and fraction prepared processing crude extract of common bean seed in respect of its
by ultrafiltration in turbidity removal were lower than those for simplicity in comparison to multistep isolation and purification
natural coagulants from the most efficient M. oleifera and other procedures.
plant materials (Ndabigengensere and Narasiah, 1998; Diaz et al.,
1999; Okuda et al., 2001; Sanchez-Martin et al., 2010). However,
Acknowledgment
this lower efficiency can be also perceived from the aspect that
it was evaluated at low water turbidity and considering that M.
The financial support from Ministry of Education and Science,
oleifera is also not efficient coagulant for low-turbidity water (Lea,
Republic of Serbia (Project No. 31002) is greatly acknowledged.
2010).
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