Bioresource Technology 97 (2006) 1455–1460

Effects of storage conditions of Moringa oleifera seeds

on its performance in coagulation
S. Katayon a, M.J. Megat Mohd Noor a,*, M. Asma a, L.A. Abdul Ghani a,
A.M. Thamer a, I. Azni a, J. Ahmad b, B.C. Khor c, A.M. Suleyman d
a
Faculty of Engineering, Universiti Putra Malaysia, 43400 Serdang, Selangor, Malaysia
b
Kolej Universiti Sains and Teknologi Malaysia, 21030 Kuala Terengganu, Malaysia
c
Indah Water Konsortium Sdn. Bhd., 50400 Kuala Lumpur, Malaysia
d
Universiti Islam Antarabangsa, 53100 Selangor, Malaysia

Received 12 May 2004; received in revised form 11 July 2005; accepted 14 July 2005
Available online 4 October 2005

Abstract

Moringa oleifera is a plant whose seeds have coagulation properties for treating water and wastewater. In this study the coag-
ulation efficiency of Moringa oleifera kept in different storage conditions were studied. The Moringa oleifera seeds were stored at
different conditions and durations; open container and closed container at room temperature (28 °C) and refrigerator (3 °C) for
durations of 1, 3 and 5 months. Comparison between turbidity removal efficiency of Moringa oleifera kept in refrigerator and room
temperature revealed that there was no significant difference between them. The Moringa oleifera kept in refrigerator and room tem-
perature for one month showed higher turbidity removal efficiency, compared to those kept for 3 and 5 months, at both containers.
The coagulation efficiency of Moringa oleifera was found to be dependent on initial turbidity of water samples. Highest turbidity
removals were obtained for water with very high initial turbidity. In summary coagulation efficiency of Moringa oleifera was found
independent of storage temperature and container, however coagulation efficiency of Moringa oleifera decreased as storage duration
increased. In addition, Moringa oleifera can be used as a potential coagulant especially for very high turbidity water.
Ó 2005 Elsevier Ltd. All rights reserved.

Keywords: Water treatment; Natural coagulant; Coagulant storage; Coagulation efficiency, Moringa oleifera

1. Introduction as AlzheimerÕs disease associated with residual alumin-

Turbidity removal is one of the important steps in a
water treatment process, which is generally achieved
using coagulants. Many coagulants are widely used in
conventional water treatment processes, based on their
chemical characteristics. These coagulants are classified
into inorganic, synthetic organic polymer and natural
coagulants. The two most commonly used primary
coagulants are aluminium and iron (III) salts (Okuda
et al., 1999). However, recent studies have pointed out
several serious drawbacks of using aluminium salts, such
*
Corresponding author.
ium in treated water and production of large sludge vol-
umes (Ndbigengesere and Narasiah, 1998). There is also
the problem of reaction of alum with natural alkalinity
present in the water leading to a reduction of pH and
low efficiency in coagulation in cold water (Haarhoff
and Cleasby, 1988). In addition, the use of alum and
iron salts is inappropriate in some developing countries
because of the high cost and low availability of chemical
coagulants (Schultz and Okun, 1983).
To ease the problems associated with chemical coag-
ulants, several studies have pointed out the introduction
of natural coagulants produced or extracted from
microorganisms, animals, or plants (Kawamura, 1991;

0960-8524/$ - see front matter Ó 2005 Elsevier Ltd. All rights reserved.
doi:10.1016/j.biortech.2005.07.031
1456 S. Katayon et al. / Bioresource Technology 97 (2006) 1455–1460
Lee et al., 1995; Ganjidoust et al., 1997). Among plant
materials that have been tested over the years, the seeds
from Moringa oleifeira have been shown to be one of the
most effective primary coagulants for water treatment.
Moringa oleifera is a tropical plant belonging to the fam-
ily of Moringaceae. Moringa oleifera is a non toxic (Gra-
bow et al., 1985) and natural organic polymer. The tree
is generally known in the developing world as a vege-
table, a medicinal plant and a source of vegetable oil
(Morton, 1991). The leaves and young seed of Moringa
oleifera are rich in calcium, iron and vitamin C (Morton,
1991), which serve as nutritious source for communities.
The fruits, which are called ‘‘pods’’ and roots of the tree,
are used as vegetables. The fruits range usually 20–
30 cm long. Each fruit contain 20 seeds. Seeds are
globular, on average weight approximately 3.0–4.0 g,
1.0–1.40 cm long and 1.0–1.7 cm wide (Goh, 2005).
According to Goh (2005), the cultivation cost for pro-
ducing 1 kg (3400 seeds) of Moringa oleifera is approxi-
mately US$2. Although cost of Moringa oleifera seems
more expensive than alum (1 kg: approximately US$1),
but is more beneficiary to communities in terms of
health and economy. Community could gain income
from the sale of the seeds to companies or institutions
processing them to produce coagulant or oil.
The active ingredients in aqueous extracts are dimeric
proteins with a molecular weight of about 13 kDa and
iso-electric point of between 10 and 11 (Muyibi and Evi-
son, 1995, 1996). Amino acid analysis and sequencing of
Moringa oleifera showed high contents of glutamine,
arginine and proline as well as total of other 60 residues
(Gassenschmidt et al., 1995). Ndbigengesere et al. (1995)
studied the efficiency and properties of Moringa oleifera
as a natural coagulant and its mechanism of coagulation
on turbid water. They discovered that the active agents
in aqueous Moringa oleifera are more effective coagu-
lants than alum. Ion-exchange columnÕs test further
showed that positive charges were a prerequisite for
coagulation to be initiated. The zeta-potential measure-
ments also indicated that the predominant mechanism
of the coagulation with Moringa oleifera appeared to
be adsorption and charge neutralization (Ndbigengesere
et al., 1995). They also revealed that Moringa oleifera
can be either used in shelled or non-shelled dry forms
seeds, however, shelled seeds are more effective (Ndbig-
engesere et al., 1995). Furthermore, sludge produced by
Moringa oleifera during coagulation is not only innocu-
ous but also four to five times less in volume than the
chemical sludge produced by alum coagulation (Ndbig-
engesere et al., 1995).
Previous researchers documented 80–99% turbidity
removal by Moringa oleifera as primary coagulant both
for raw waters and synthetics turbid waters (Muyibi and
Okufu, 1995; Ndbigengesere et al., 1995; Muyibi and
Evison, 1996). However, Muyibi and Okufu (1995)
found that Moringa oleifera might not be an efficient
coagulant for low turbid water. They documented that
the residual turbidity of samples increased with the de-
crease in initial turbidity at optimum dosage of Moringa
oleifera. They achieved only 50% turbidity removal from
low turbidity surface waters (23–90 NTU). Okuda et al.
(1999) concluded that use of Moringa oleifera for drink-
ing water treatment may not be appropriate since tur-
bidity of raw water for drinking water is usually low.
Although there are many studies been carried out on
Moringa oleiferaÕs efficiency as a coagulant (Muyibi and
Okufu, 1995; Muyibi and Evison, 1995, 1996), studies
on the effects of storage condition on its performance
have not been established. Thus present study is aimed
to examine the effects of storage condition on the perfor-
mance of Moringa oleifera in coagulation.
2. Methods
2.1. Preparation of Moringa oleifera seeds powder
The Moringa oleifera was obtained from Klang,
Selangor area. Good quality seeds were identified from
those, which were not rotten, old, infected with diseases,
brownish and dried once opened. The seeds were dried
in the oven (Memmert, ULE 400, Germany) for 24 h
at 50 °C. A rice husk-removing machine (Satake, THU
class) was used to remove the hulls and wings from
the kernels. The kernels were crushed and ground to a
medium fine powder with a domestic food blender
(Moulinex, France).
2.2. Preparation of Moringa oleifera seeds extract
A 5000 mg of Moringa oleifera seeds powder was
placed in a beaker containing 0.2 l of distilled water.
The mixture was blended using domestic blender (Moul-
inex) for 2 min at high speed to extract the active in-
gredient of Moringa oleifera. The suspension was then
filtered through a muslin cloth in a beaker and the fil-
trate made up to 0.5 l to give a stock solution of
10,000 mg/l. 10,000 mg/l of Moringa oleifera stock solu-
tion was used for jar test trials that were conducted
to determine optimum dosages of Moringa oleifera on
water samples of varying initial turbidities.
2.3. Preparation of water samples
Synthetic turbid water samples were prepared by add-
ing kaolin (Laguna Clay, California, USA) into distilled
water. Ten grams of kaolin was added to 1 l of distilled
water. The suspension was stirred slowly at 20 rpm for
1 h in a jar test apparatus (BIBBY Stuart Scientific,
UK) for uniform dispersion of kaolin particle. The sus-
pension was then allowed to stand for 24 h to allow for
complete hydration of the kaolin. The maximum parti-
 S. Katayon et al. / Bioresource Technology 97 (2006) 1455–1460 1457

cle size remaining in the kaolin suspension was esti-
mated by Scanning Electron Microscopy (JEOL 6400,
Tokyo, Japan) to be around 1 lm. This kaolin suspen-
sion was used as the stock solution for the preparation
of water samples of varying turbidities for the coagula-
tion tests. Three types of turbidities were carried out
(Muyibi and Evison, 1995, 1996) namely; medium tur-
bidity (50–100 NTU), high turbidity (100–200 NTU),
and very high turbidity (greater than 300 NTU).
2.4. Packing and storage of Moringa oleifera
The ground Moringa oleifera seeds were grouped into
two and packed in different types of packages namely;
closed container and open container. Closed container
used in this study was a glass bottle with plastic cap
and open container was 500 ml glass beaker. The pack-
ages were stored at two different temperatures namely;
room temperature (28 °C) and refrigerated (3 °C). The
storage durations were for 1 month, 3 months and
5 months.
of the synthetic water, and placed on the floc illuminator
and agitated at the preselected intensity of rapid mixing
(Table 1). During rapid mixing, the coagulant dosage of
Moringa oleifera was added into each beaker simulta-
neously using 20 ml test tubes. After rapid mixing, the
preselected intensity of slow mixing was quickly estab-
lished and the beakers were then carefully removed from
the floc illuminator and left for the sedimentation phase.
After settling, 20 ml of the sample was taken from the
middle of each beaker for turbidity measurement. For
determination of optimum dosage, different amount of
Moringa oleifera stock solution were added into the
beakers and the amount that gave the lowest turbidity
was the optimum dosage for that particular water.
Secondly, randomized experimental runs with optimum
Moringa oleifera dosages kept in varied storage condi-
tions in terms of packing and temperature were carried
out as shown in Table 2. Turbidity measurements were
conducted using Turbidimeter (HACH, 21200P). pH
values of samples were measured using pH meter
(CyberScan pH 2000).

2.5. Coagulation test 2.7. Statistical analyses

Coagulation test was carried out using jar test
(BIBBY Stuart Scientific, UK). The study involved
rapid mixing, slow mixing and sedimentation in a batch
process. Six glass beakers of 500 ml capacity were filled
with the water samples and agitated simultaneously, and
the rotational speed were varied accordingly, allowing
simulation of different mixing intensities and resulting
flocculation process.
2.6. Experimental runs
The SPSS statistical package (Version 11.0) was used
for all statistical analysis. All statistical significance was
considered when p < 0.05. One-way analysis of variance
(ANOVA), with TukeyÕ HSD test was carried out to
verify the significance of differences among the means.
Independent sample test (t-test) was used to confirm
the significant differences between the two means. Cor-
relation between two variables was analyzed using
bivariate analysis of variance. All experiments done in
triplicates.

First, sets of experimental runs were carried out for

optimization of Moringa oleifera dosages for coagula- 3. Results and discussion
tion of water samples. Beakers were filled with 500 ml
3.1. Optimization of Moringa oleifera dosages

Table 1 Results on optimization of Moringa oleifera dosages

Operating variables used to run the jar test (Muyibi et al., 2002a,b)
for medium, high and very high turbid water are shown
Rapid mix Rapid mix Slow mix Slow mix Settling in Figs. 1–3, respectively. Summarized data presented in
velocity duration velocity duration time
Table 3, revealed that the optimum dosage of Moringa
(rpm) (min) (rpm) (min) (min)
oleifera seeds that gave the lowest residual turbidity
100 4 40 25 30
(p < 0.05) was dependent on the initial turbidity. As

Table 2
Experimental runs to evaluate coagulation performance of Moringa oleifera kept in different durations and conditions
Storage conditions Storage conditions (temperature)
(packing) Room temperature (28 °C) Refrigerator (3 °C)
a
Initial turbidity of water samples (NTU)
Open container MT (80–90) HT (185–193) VHT (385–395) MT (88–89) HT (185–187) VHT (385–390)
Closed container MT (87–89) HT (185–187) VHT (385–390) MT (88–89) HT (185–193) VHT (385–390)
a
MT: medium turbidity, HT: high turbidity, VHT: very high turbidity.
1458 S. Katayon et al. / Bioresource Technology 97 (2006) 1455–1460

90 36

80 34
Residual turbidity (NTU)

Residual turbidity (NTU)

70
32

60
30
50
28
40

26
30

24
20

10 22
100 120 140 160 180 200 360 380 400 420 440 460
Dosage (mg/l) Dosage (mg/l)

Fig. 1. Optimization dosage of Moringa oleifera for medium turbidity Fig. 3. Optimization dosage of Moringa oleifera for very high turbidity
water sample. water sample.

80 Table 3
Optimum dosages of Moringa oleifera for different water samples
70 Sample Moringa oleifera Turbidity Turbidity
concentration (NTU) removal (%)
Residual turbidity (NTU)

60 (mg l 1)
Initial Final
Medium 160 87.8 ± 2.1 21.3 ± 1.7 79 ± 2.0
50 turbidity
High 300 194 ± 3.8 22.0 ± 2.2 89 ± 3.2
40 turbidity
Very high 400 390 ± 4.5 23.6 ± 3.6 94 ± 4.1
30
turbidity

20
10
260 280 300 320 340
Dosage (mg/l)
Fig. 2. Optimization dosage of Moringa oleifera for high turbidity
water sample.
initial turbidity of water sample was increased, the re-
quired optimum dosage of coagulant also increased
(r2 = 0.985). For high and very high turbidities, the opti-
mum dosage of Moringa oleifera were 300 mg l 1 and
400 mg l 1, respectively, while for medium water
160 mg l 1 produced highest turbidity removal (signifi-
cantly, p < 0.05). Increasing dosage of coagulant did
not improve the removal of turbidity, in fact this
increased significantly (p < 0.05) the residual turbidity
of the coagulated sample. This overdosing resulted in
the saturation of the polymer bridge sites and caused
restabilization of the destabilized particles due to insuf-
ficient number of particles to form more interparticle
bridges (Muyibi and Evison, 1995, 1996).
The optimum dosages found in this study were signif-
icantly (p < 0.05) higher than those obtained by previous
360 researchers (Muyibi and Evison, 1995). They docu-
mented the optimum dosages of about 100 mg l 1 and
50 mg l 1for moderate to high turbidities (250–550
NTU) and low to moderate turbidities (50–150 NTU),
respectively. This difference might due to difference in
experimental conditions employed, such as settling time,
mixing velocity gradient and mixing durations. Compar-
ison between optimum dosage obtained in this study
and those obtained by Muyibi and Evison (1996) also
revealed insignificant differences (p < 0.05), which might
due to usage of different species of Moringa oleifera
(Jahn, 1988).
3.2. Effect of storage durations and conditions of
Moringa oleifera seeds on its efficiency
The Moringa oleifera seeds were kept in open con-
tainer at room temperature, closed container at room
temperature, open container in refrigerator and closed
container in refrigerator for 1, 3 and 5 months. Residual
 S. Katayon et al. / Bioresource Technology 97 (2006) 1455–1460 1459

turbidity of coagulated water using Moringa oleifera 400

stored at different conditions and durations are mea-
sured and results are presented in Figs. 4–7.
In general, for all experimental conditions, turbidity

Residual turbidity (NTU)

300
removal efficiency of Moringa oleifera increased as
initial turbidity of water sample increased. The highest
turbidity removal efficiencies were recorded for water
200 Type of water
sample with very high initial turbidity. This result is
same with those reported by Muyibi and Evison
Medium turbid
(1995). They have also documented that at the optimum
dosage of Moringa oleifera residual turbidities decreased 100
and removals increased with increasing initial turbidity. High turbid

Turbidity removal up to 98.5% was recorded for a water

sample with high initial turbidity of 600 NTU (Muyibi 0
Very high turbid

and Evison, 1995). Increase in suspended particles avail- Fresh 1 3 5

Storage durations (Months)

Fig. 6. Residual turbidity of water coagulated using Moringa oleifera

500
kept in open container in refrigerator.
450

400
Residual turbidity (NTU)

400
350

300

Type of water
Residual turbidity (NTU)

250 300
200
Medium turbid
150
200 Type of water
100 High turbid

50 Medium turbid
Very high turbid
0 100
Fresh 1 3 5
High turbid
Storage durations (Months)

Fig. 4. Residual turbidity of water coagulated using Moringa oleifera Very high turbid
0
kept in open container at room temperature. Fresh 1 3 5
Storage durations (Months)

Fig. 7. Residual turbidity of water coagulated using Moringa oleifera

450 kept in closed container in refrigerator.

400

350
able for adsorption and inter-particle bridge formation
Residual turbidity (NTU)

300 in water sample with higher initial turbidity may con-

tribute in higher turbidity removal efficiency (Birkner
250
and Morgan, 1968).
Type of water
200 Comparison between residual turbidity of water trea-
ted with Moringa oleifera kept for different durations re-
150 Medium turbid
vealed that an increase in storage duration of Moringa
100 oleifera from 1 to 5 months increased the residual tur-
High turbid
bidity in most of the experimental conditions. The Mor-
50
inga oleifera kept for 1 month at all conditions showed
Very high turbid
0 the best performance in terms of turbidity removal.
Fresh 1 3 5 Exception was in the case of Moringa oleifera which
Storage durations (Months)
was kept in refrigerator in open container and applied
Fig. 5. Residual turbidity of water coagulated using Moringa oleifera on very high turbidity water sample (Fig. 6). In this case
kept in closed container at room temperature. the difference between residual turbidity of treated water
1460 S. Katayon et al. / Bioresource Technology 97 (2006) 1455–1460
samples with Moringa oleifera stored for different dura-
tions was insignificant (p < 0.05).
As presented in Figs. 4 and 5, residual turbidity of
treated water with Moringa oleifera kept at room tem-
perature in open container and closed container found
to be in the same range. Results revealed that there
was no significant (p < 0.05) difference between residual
turbidity of treated water with Moringa oleifera kept at
open or closed container in room temperature. The
residual turbidity of treated water with Moringa oleifera
kept in closed container at room temperature were
found as 60–90%, 78–92% and 90–98% for medium,
high and very high turbidity water samples, respectively,
whereas for those kept in open container were 65–92%,
73–92% and 97–98% for medium, high and very high
turbidity water samples, respectively.
Comparison between residual turbidity of treated
water with Moringa oleifera kept in closed and open
container in refrigerator showed that there were no sig-
nificant (p < 0.05) differences between them (Figs. 6 and
7). The Moringa oleifera kept in closed container in
refrigerator achieved 70–83%, 85–88% and 97–98%
turbidity removal efficiency for medium, high and very
high turbidity water, respectively. These values are in
the same range with those obtained for Moringa oleifera
which kept in open container in refrigerator.
Biological product such as Moringa oleifera will be
affected as all biological materials to humidity and
temperature. Therefore, storage condition is an impact
consideration. Presence of moisture content within the
biomateials which often causes the deterioration of
biomaterials warrant further study.
4. Conclusions
The Moringa oleifera seeds were found as a potential
coagulant, especially for pretreatment of very high tur-
bidity water. The Moringa oleifera kept in refrigerator
and room temperature for one month showed better
performance in terms of turbidity removal than those
kept for 3 and 5 months, at both containers. However,
the difference between turbidity removal efficiency of
Moringa oleifera kept in refrigerator and room temper-
ature was insignificant. This provide positive input to
explore further on preservation of Moringa oleifera in
order to commercialize the product.
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