~

War. Res. Vol. 29, No. 2, pp. 703-710, 1995

Pergamon

0043-1354(94)00161-8

Copyright 1995 Elsevier Science Ltd

Printed in Great Britain. All rights reserved
0043-1354/95 $7.00 + 0.00

ACTIVE AGENTS A N D MECHANISM OF COAGULATION

OF T U R B I D WATERS USING M O R I N G A OLEIFERA
ANSELME NDABIGENGESEREI, K. SUBBA NARASIAH 1 . ~ and BRIAN G. TALBOT2
tDepartment of Civil Engineering and 2Department of Biology, Universit6 de Sherbrooke, Sherbrooke,
Quebec, Canada J1K 2RI

(First received June 1993; accepted in revised form May 1994,)

Abstract--Moringa oleifera is a tropical plant whose seeds contain an edible oil and water soluble
substance which has excellent coagulation properties for treating water and wastewater. The et~ciency and
properties of Moringa oleifera as a natural coagulant in water treatment were studied and compared with
alum, which is presently the most widely used industrial coagulant. It is shown that the active agents in
aqueous Moringa extracts are dimeric cationic proteins, having molecular weight of 13 kDa and isoelectric
points between 10 and I 1. The mechanism of coagulation with Moringa oleifera appears to consist of
adsorption and neutralization of the colloidal charges. Compared to alum, the optimal dosage of shelled
Moringa oleifera seeds was almost the same (50 mg/1). In case of the non-shelled seeds, the dosage is greater
(500 mg/l) for low initial turbidity waters. The purified proteins are more effective coagulants than alum.
As a coagulant, Moringa is non-toxic and biodegradable. It is environmentally friendly, and unlike alum,
does not significantly affect the pH and conductivity of the water after the treatment. Sludge produced
by coagulation with Moringa is not only innocuous but also four to five times less in volume than the
chemical sludge produced by alum coagulation. So, as a coagulant, Moringa oleifera may be a potentially
viable substitute to alum.

Key words--natural coagulants, Moringa oleifera, cationic proteins, coagulation-flocculation, water

treatment

INTRODUCTION
Aluminum salts are the most common synthetic
coagulants used in water and wastewater treatment
all over the world (Degremont, 1989; Bratby, 1980).
However, recent studies by several workers (Mallevialle et al., 1984; Miller et al., 1984; Letterman and
Driscoll, 1988) have raised doubts about the advisability of introducing aluminum into the environment. Ferric salts and synthetic polymers have been
used as alternatives but with limited success due to
the fact that their impact on living beings is not fully
investigated (Letterman and Pero, 1990; Goppers and
Straub, 1976; Aizawa et al., 1990). Besides, many
developing countries can hardly afford the costs
of imported chemicals for water and wastewater
treatment.
Natural coagulants of vegetable and mineral origin
were in use in water and wastewater treatment before
the advent of synthetic chemicals like aluminum and
ferric salts. Previous studies however, have not determined whether such natural coagulants are economically and environmentally more acceptable than
chemical coagulants (Hespanhol and Selleck, 1975;
Jahn, 1986). Since recently there has been more
interest in the subject of natural coagulants, especially to alleviate problems of water and waste*Author to whom all correspondence should be addressed.
703

water treatment in developing countries (Jahn, 1981)

and to reuse some by-products (Kawamura, 1991a).
Moringa is a tropical plant belonging to the family
of Moringaceae. According to Jahn (1988) 14 species
have so far been identified and all possess coagulant
properties in varying degrees. Moringa oleifera is the
most widespread species which grows quickly, even
on medium soils having relatively low humidity
(Jahn, 1991).
Earlier studies have found the plant to be non-toxic
(Grabow et al., 1985), and recommended its use as
coagulant in developing countries (Jahn, 1981, 1986,
1988; Barth et al., 1982; M/iller, 1980; Olsen, 1987;
Bhole, 1987). Encouraged by results of these studies,
many developing countries have turned to using this
plant as a viable coagulant in water and wastewater
treatment on a small scale (Karerwa, 1986; Kaiser,
1989; Ndikumana, 1987; Ndabigengesere, 1988;
Sutherland et aL, 1989; Sekamana, 1989, Nimubona,
1990).
Preliminary studies on the active ingredients of
Moringa oleifera as a coagulant have suggested that
the active components are cationic peptides of molecular weight ranging from 6 to 16 kDa and isoelectric pH value of 10 (Fink, 1984; Gassen et al., 1990;
Gassenschmidt et al., 1991).
The coagulating property of Moringa oleifera has
already been proved, but its mechanism of reaction is
not yet fully expanded. The main objectives of the

ANSELMENDABIGENGESEREet a/,

704

present study are to purify and characterize the active

c o m p o n e n t s in order to investigate their m e c h a n i s m
of coagulation a n d to c o m p a r e Moringa as a coagulant with alum. In order to explain the m e c h a n i s m of
coagulation with Moringa oleifera, we refer to the
four basic m e c h a n i s m s of classic coagulation theory,
namely, compression of the double layer, a d s o r p t i o n
a n d neutralization o f charges, a d s o r p t i o n a n d bridging, a n d then e n m e s h m e n t in the precipitate (Weber,
1972; Benefield et al., 1982; Edzwald et al., 1989;
Dentel a n d Gosset, 1988; Attia, 1987).
MATERIALS AND METHODS

Coagulants used
The Moringa oleifera used as a coagulant in this study
comes from Burundi in Central Africa. The green and dry
pods of Moringa were picked for the present study in June
1991 and transported to the Environmental Engineering
Laboratory of the Universit6 de Sherbrooke, Canada, where
they were stored until used.
The aluminum sulphate [AI2(SO4)3.18H20 ] used in this
study was of laboratory grade (Anachemia AC-405).

Preparation of Moringa
In this study, the green and dry pods, the shelled and
non-shelled seeds, and also the bark enveloping the seeds
were tested for their coagulation potential. The extraction of
the active ingredients was carried out as follows: the raw
material was ground to powder in a domestic blender, the
powder was added, in the appropriate proportion, to the
solvent under investigation, and the whole mixture was
stirred for 30 min. The suspension was then filtered firstly
through Whatman No. 42 paper and then through a
0.45 t~m nylon membrane.
Different solvents were tested for their capacity to extract
the active ingredients, namely, water, petroleum ether,
acetone, chloroform and hexane. Only water was able to
extract the active agents in the coagulation with Moringa.
Successive extractions were also conducted to study the
compatibility of the exploitation of vegetable oil and the
active agents in the coagulation process. After a first series
of tests, water was retained as solvent and the standard
concentration of 5% by weight (crude powder/solvent) was
adopted, since higher concentrations were too hard to filter.
Solids obtained by lyophilisation and by evaporation at
20C and also at 105C, of Moringa solutions were also tried
for their coagulating activity.

"'Synthetic" turbid water

For the purpose of our experiments, a model turbid water
was prepared by adding kaolin to tap water. Five grams of
laboratory grade kaolin (Anachemia, AC-5302) were added
to a litre of water. The suspension was stirred for 30 min and
then allowed to settle for 24hs. The supernatant was
carefully removed and stored in a plastic bottle. The maximum particle size remaining in the kaolin suspension was
estimated to be about 2/~m by Stokes law. The kaolin
suspension was diluted using tap water to obtain any desired
turbidity. For example, the comparative tests of Moringa
and alum were conducted on the turbid water prepared by
diluting 400 ml of the kaolin suspension in 201 of tap water,
resulting in an initial turbidity of 105 NTU. This model
water does not represent real water in any country, but is
a stable suspension used here to study the mechanism of
coagulation.

Coagulation test
The jar test has been and is still the method widely used
to evaluate coagulation-flocculation processes (Hudson,
1981; Kawamura, 1991b). Coagulation activity of each

Moringa extract was verified by the jar test. Coagulation

tests were conducted using the jar test equipment of Phipps
& Bird having a base floc illuminator. Six beakers were filled
with 1 1 of the suspension to be tested and were agitated
simultaneously at a speed varying from 0 to 100 rpm. It may
be added here that there is no standard method for conducting the jar test. Hence we adapted the test to suit our needs:
a rapid mix at 100 rpm for 2 min followed by a slow mix of
20 min at 40 rpm. This was followed by 30 min of sedimentation. One litre of the synthetic water was used for the test
in each of the 6 beakers placed on the base illuminator. The
speed of rotation was adjusted to 100 rpm and the content
of these beakers was agitated while the required amount of
the coagulant was added to the beakers. A chronometer was
then started to control the 2min at 100rpm for rapid
mixing, the 20 min at 40 rpm for slow mixing and the 30 min
of sedimentation.
Comparative coagulation tests were run under the same
conditions as described above but using 5% alum solution
instead of Moringa.
Chemical analysis on Moringa
Various tests were performed on Moringa oleifera in its
various forms, the principal objective being the determination of its general composition and its potential use as a
natural coagulant in water and wastewater treatment.
Characterization was done by conducting elemental
analysis of the seeds, that is the content of carbon, nitrogen,
hydrogen and oxygen. Also determined were the protein, fat
and sugar contents. The elemental analysis was conducted
on the powdered shelled Moringa seeds as well as on
non-shelled Moringa seeds using a Perkin-Elmer elemental
analyzer (model 240C). The protein content, sugar and fat
of both the powder and the 5% solution of shelled and
non-shelled Moringa seeds were conducted by the AgriAnalysis laboratory at Lennoxville, Quebec, using standard
methods.

Purification and characterization of active agents

Active agents were purified and characterized using different techniques, namely: dialysis, ultrafiltration, lyophilisation, ion-exchange, chemical precipitation and electrophoresis. These techniques are adequately described in
numerous references (Jirgensons, 1962; Franks, 1988;
Scopes, 1987) and therefore will not be elaborated here.
Coagulation activity of each fraction was tested as already
described by the jar test.
Dialysis tubes (Spectra/Por) with molecular weight cutoff
from 6000 to 8000Da, and from 12,000 to 14,000Da
(Spectrum Medical Industries) were used.
Ultrafiltration was carried out using a Millipore cell (GS
Amicon) and the membranes YM2, YMI0 and YM30,
which are permeable to compounds having respectively
molecular weights less than 1000, 10,000 and 30,000 Da.
Lyophilisation of various fractions of Moringa solutions
was carried out using a Labconco freeze dryer (model 4.5).
Precipitation of active proteins from Moringa solutions
was carried out using 80-100% saturated solutions of
ammonium sulphate (Anachemia, ACS 093). The required
amount of salt was added directly to the Moringa solutions.
Following homogenization for 30 min using a magnetic
stirrer, the precipitate was separated by filtration on a
membrane filter (0.45 #m) and then redissolved in distilled
water or in 0.05 M phosphate buffer (pH of 7.0).
In order to determine the nature of the charges carried by
the coagulant active proteins, a cation-exchange resin,
namely carboxymethyl cellulose (CM cellulose C-50), and
an anion-exchange resin, namely diethylaminoethyl (DEAE
Sephadex A-50) were employed. The ion exchange characteristics were analyzed using 30 cm x 1 cm columns of either
DEAE Sephadex of CM cellulose (Pharmacia) equilibrated
either in 0.05 M Tris-HCl buffer (pH of 7.4) or in 0.05 M
phosphate buffer (pH of 7.0), or simply in distilled water.

Coagulation of turbid waters

Table I. Coagulation activity of different Moringaforms
Forms of Moringa
Coagulation activity
Green pods:
--Whole pods
absent
--Seeds
absent
--Bark of green pods
absent
--Dried green pods
absent
Dried pods:
--Whole pods
--Non-shelled seeds
Unfiltered
--Filtered
--Residual solids
--Shelled seeds
Unfiltered
--Filtered
--Residual solids
Bark of pods
Bark of seeds

absent
present
present
absent
present
present
absent
absent
absent

The columns were eluted, if necessary, with a concentration

gradient 0-1 M NaCI in the same buffers.
Studies on the purified proteins were carried out using an
S-18 reverse phase column on a Beckman HPLC system.
The purity and molecular weights of the proteins were
analyzed by sodium dodecylammonium sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) under reducing
and non-reducing conditions using 15% gels. The results
were confirmed by molecular sieving in the presence or
absence of 1 mM mercaptoethanol on a 50cm x lcm
column of Sephadex G-100 (Pharmacia) in 0.05 M phosphate buffer (pH of 7.0). Purified active extracts were
desalted against water, lyophilised and then stored at ambient temperature.

Parameters analyzed
The following parameters were analyzed in the raw and
treated water using Standard Methods (APHA, 1992): pH
value, turbidity, conductivity, sludge volume and zeta potential. Turbidity measurements were conducted using a
Hellige turbidimeter (series 12520). The pH value was
measured with a Hach-one pH meter. Conductivity was
measured also with the same brand name instrument. Sludge
volume was measured using Imhoff cones. Zeta potential
was measure with a zetameter Zetasizer II of Malvern. The
objective of the zeta potential measurements was to determine the mechanism of coagulation which depends upon the
electrostatic forces between charges carried by the colloidal
particles.
RESULTS AND DISCUSSION

plant. The extraction in water of the whole dry pods

did not give any positive result. Therefore, for
Moringa to serve as a coagulant one cannot directly
pulverize the dry pods. If the dry seeds are removed
and ground separately, the coagulation activity was
found in the seeds. Similarly, the filtered and
unfiltered solution of Moringa seeds has coagulant
properties, whereas the residual solids do not show
any sign of coagulation activity.
The conclusion from the foregoing results is that
the active agents in the coagulation with Moringa are
water soluble materials. Coagulation activity is present in shelled and in non-shelled seeds but only after
the pods are dry. From Table 1, it is also to be noted
that the bark around the seed has no coagulation
properties. Comparing the shelled and non-shelled
Moringa seeds, we found that the latter was almost as
good as the former as a coagulant, for an initial
turbidity of 426 N T U . When the initial turbidity was
lowered to 1 0 5 N T U however, shelled seeds were
more effective coagulant then non-shelled ones, as
will be discussed later. This means that when treating
highly turbid waters, it is not necessary to separate
the shell from the seed, before using Moringa seeds as
a coagulant. Removing shells from Moringa seeds is
a tedious process consuming time and expense.
Figure 1 shows the results of coagulation of a
model water having an initial turbidity of 426 N T U ,
with 5% solutions of shelled and non-shelled
Moringa seeds. On an industrial scale, for highly
turbid waters, it is advantageous to use the nonshelled seeds in order to save the labour and energy
required to separate one from the other.
We also studied the effect of the concentration of
Moringa solution on the coagulation activity. Figure
2 shows turbidity removal after coagulation with
different concentrations of non-shelled Moringa seeds
solutions. More concentrated Moringa solutions are
better because small volumes are required to get the
optimal dosage. The optimal dosage was 500 mg/1 for
500 --

Forms of Moringa and coagulation activity

Table 1 sums up the results of coagulation activity
of various forms of Moringa. This is a qualitative
assessment of the coagulating capacity of Moringa
pods and seeds. During the experiment, coagulant
property was manifested by formation of visible flocs,
easily recognized by naked eye following flow agitation of the turbid water. Considerable turbidity
(80-90%) was removed after sedimentation in the jar
test. In this case the coagulation activity was qualified
as "present". On the contrary, if the turbidity removal was below 30% the result was described as
"absent". As can be seen from the Table 1, all
preparations based on green pods of Moringa do not
possess any coagulation activity. This means that for
exploitation of Moringa seeds as a coagulant, the
pods should be allowed to dry completely on the

705

~" 400
[-Z
300

* Shelled Moringa seeds (SMS)

O Non-shelled Moringa seeds (NSMS)

200

I00

10

20

30

Dosage (mill)
Fig. 1. Coagulating activity of shelled (SMS) and nonshelled (NSMS) Moringa seeds

ANSELME NDABIGENGESERE el al.

706
120

Table 3. Percentage (by mass) of proteins, lipids and sugars in

Moringa seeds

i~.

Moringa seeds
preparation

* 0.5%

"N

50

100

150

Dosage (mill)
Fig. 2. Coagulating activity of non-shelled Moringaseeds at
different concentration in aqueous solution
all the four concentrations studied. The 5% Moringa
solution was retained as the standard in this work.
The coagulant dosage expressed in mg/l refers to the
mass of the crude powder used to prepare the
Moringa solution, but after filtration more than 80%
stayed in solid state. This is why the coagulant dosage
is expressed mostly in ml/1 throughout this study.

General composition of Moringa oleifera

Data presented in this section are simple averages
of at least two analyses.
Results of elemental analysis of shelled and nonshelled Moringa seeds are presented in Table 2.
Organic matter consists of the six principal elements,
namely carbon (C), oxygen (O), hydrogen (H), nitrogen (N), phosphorus (P) and sulphur (S). The principal groups of the above constituents are proteins,
carbohydrates, lipids, lignins and free amino acids
(Jirgensons, 1962). In our studies, we found that the
shelled Moringa seeds contain 55% carbon, 8.5%
hydrogen and 6% nitrogen. The remaining 31%
consists of oxygen and trace elements. As can be seen
from Table 2, the non-shelled Moringa seeds trail
closely the shelled Moringa seeds in all the elements
analyzed, all of them being inferior by a small margin
however. Nitrogen is a common component in all
proteins and free amino acids (Rogers, 1991). Some
authors (Strong, 1974) use the proportionality factor
of 6.25 to estimate the protein content of organic
matter from the available values of nitrogen.
The mass of the seed cotyledon is 70% of the total
mass of non-shelled Moringa seeds and the remaining
30% is for the shell. Using the hypothesis that the

Proteins
(%)

Lipids
(%)

Carbohydrates
(%)

Shelled
--Powder
--Solution
--Residual solids

36.7
0.9
29.3

34.6
0.8
50.3

5.0
-1.3

Non-sheUed
--Powder
--Solution
--Residual solids

27.1
0.3
26.4

2 I. 1
0.4
27.3

5.5
---

nitrogen is contained only in the cotyledon, the

nitrogen content of the whole non-shelled Moringa
seeds can be estimated from Table 2 as follows:
(100"6.1 + 70"5)/(2"100) = 4.8%, which is very close
to the measured nitrogen content of 5% for nonshelled seeds. It appears that almost all the proteins
and amino acids are in the seed cotyledons.
Table 3 shows the results of the analysis of Moringa
oleifera into proteins, lipids and carbohydrates. The
shelled Moringa seeds contain about 37% proteins,
27% in the non-shelled ones, whereas there is close to
35% of lipids in the shelled Moringa seeds, and 21%
in the non-shelled seeds. Carbohydrates (as oligosaccharides) represent 5% of the shelled seeds whereas
they are 5.5% of the non-shelled Moringa seeds.
The carbohydrates content is very low whereas the
high lipids content explains why Moringa seeds can
be used as a source of vegetable oil. The lipids are not
soluble in water, this is why their content in the
residual solids increases to 50.3% in case of shelled
seeds and to 27.3% for non-shelled ones.
The percentage of extractable matter of Moringa
seeds using various solvents like ether, hexane, carbon tetrachloride, acetone, water and methanol,
along with their respective coagulating activity is
shown in Table 4. Only the water extract possess
coagulation activity. The extracts with petroleum
ether, hexane and chloroform are mainly vegetable
oils, whereas the extract with acetone and methanol
are mainly carbohydrates (Jirgensons, 1962). It is
clearly possible to extract oil first and then use the
aqueous extract as a coagulant. This dual exploitation is even advantageous for isolating and purifying
the active agents in the coagulation with Moringa
seeds and also for the reduction of organic matter
concentration in the treated water.

Table 4. Coagulation activity of extracts from Moringa with different

solvents

Solvent
Table 2. Elemental analysis of Moringa (% by mass)
Sample
Shelled Moringa seeds (SMS)
Non-Shelled Moringa seeds
(NSMS)

N (%)

C (%)

H (%)

6.1

54.8

8.5

5.0

53.3

7.7

Petroleum ether
Hexane
Chloroform
Acetone
Water
Methanol

Quantity of
extractable matters
(% by mass)

Coagulation
activity

33.9
27. l
26.6
24.4
25.0
26.3

absent
absent
absent
absent
present
absent

Coagulation of turbid waters

MW
kDa
9 7 ~
4 6 ~

3 1 ~

2 1 ~

14--

Std
B
C
Fig. 3. SDS PAGE separation on Moringa proteins. 10/ag
of polled coagulating proteins, purified by CM-cellulose to
the gel. (B) under reducing conditions (1 mM mercaptoethanol); (C) under non-reducing conditions. Std: molecular weight standards

Characterization of the active agents in coagulation

with Moringa
The aqueous extract of Moringa is far from being
pure. It is a solution consisting principally of proteins, lipids and carbohydrates. A series of experiments were carried out to characterize the
coagulating and flocculating agents of this mixture.
Precipitation tests with ammonium sulphate
showed that the precipitate has excellent coagulant
properties. The supernatant did not possess any
coagulation activity. Ammonium sulphate precipitates are known to be proteins (Scopes, 1987). This
result supports the previous works (Fink, 1984;
Gassen et al., 1990; Gassenschmidt et al., 1991),
suggesting that the agents responsible of the coagulation and flocculation with Moringa oleifera are
water soluble proteins. Fink (1984) proposed that the
active agents are water soluble proteins of molecular
weights between 6000 and 16,000, and he separated
two active fractions (MO1 and MO2) on columns.
Later, Gassen (1990) purified the two fractions
and found that they are proteins of 6500Da.
Gassenschmidt (1991) suggested the sequence of the
two polypeptides.
Dialysis experiments showed that coagulation activity stayed inside of the dialysis tubes with molecular weight cutoff from 12,000 to 14,000 Da.
Similarly, uttrafiltration tests showed that the active ingredients were retained on the 10 kDa membranes, but passed through the 30 kDa membranes.
W R 29'2

707

Determination of the molecular weight of the

proteins extracted from Moringa seeds by molecular
sieving method showed molecular weight of 6.5 kDa
under reducing and of 13 kDa under non-reducing
conditions. An identical result was obtained by
S D S - P A G E tests, as shown in Fig. 3. The photograph shows the migration of a series of molecular
weight standards, ranging from 14 to 2000 kDa, and
the Moringa proteins, in presence and absence of
mercaptoethanol. Together these results showed that
the native protein was a dimeric of 13 kDa with
subunits of about 6.5 kDa. The subunits are liked by
S-S bonds. In reducing conditions, our results
confirm the suggestion by Gassen et al. (1990) that
the active proteins have a molecular weight of
6.5 kDa. Both monomers and dimers retain their
coagulant properties.
Ion-exchange column experiments showed that the
active proteins carry positive charges. When 5%
Moringa solution was applied to DEAE cellulose at
pH of 7.4, the active proteins did not bind. However,
with the CM cellulose, the active proteins were bound
and could be eluted in a very pure state at 0.6-1.0 M
NaC1, as shown in Fig. 4. There were three active
peptides isolated from this column (A1, A2 and A3)
which is consistent with their being at least two
different monomers which can associate at random.
Experiments using polyacrylamide gel isoelectric
focusing in a pH gradient from 3 to 11, using the
purified Moringa proteins from the CM cellulose
column showed that these proteins had isoelectric
points between 10 and 11, thus clearly demonstrating
their highly cationic nature.
Lyophilisation of crude Moringa solution and also
the purified Moringa proteins results in a white
powder, which is soluble in water and its solution has
very good coagulant properties. The Moringa proteins are stable in this dried form and remain so, even
after 6 months of storage without any special precaution as to the pH and temperature. This means
that isolation and purification are relatively easy
0.20 --

-'] 1.2

DO 280 nm
0.15

....

GradientNaCl

// " //1

1,0 ~
0.8

o.lo

0.6
9

t~

e-

0.4

0.05
0.2
0

50

100

150

200

Elution volume (ml)

Fig. 4. Elution of Moringa proteins from a CM cellulose
column. AI, A2 and A3 represent discrete factions with
coagulating properties.

ANSELMENDABIGENGESERE et

708
120

10
, t ~ k
. . . .

./

[-.

A--"~" . . t

-t

80 ~* AAI'

Zeta

i~

* NTU

~3

- -to
-20

-30

r~

40

\
0

_
I

10

20

30

-40

-50

Dosage (ml/l)
Fig. 5. Residual turbidity and zeta potential of water as
function of dosage of non-shelled Moringa seeds used as a
coagulant
compared to the costly and laborious manipulations
for other proteins (Franks, 1988). However, slow
evaporation of Moringa solution at ambient temperature and also at about 105C resulted in some denaturation of the proteins which became difficult to
solubilize and less efficient in coagulation. Hence, for
best results in the preparation of Moringa as a
coagulant, if not freeze drying, fast spin evaporation
may be the most economically feasible approach.

Mechanism of coagulation with Moringa

As mentioned before, the active agents in coagulation with Moringa are water soluble cationic proteins. The zeta potential of a 5% solution of
non-shelled Moringa seeds was found to be + 6 mV,
confirming that even if the solution is a heterogenous
complex mixture, the positive charges are predominant in the solution.
The zeta potential of the synthetic water was
- 4 6 inV. This means that at a pH of around 7, the
kaolin particles were charged negatively. Hence coagulation of the kaolin suspension using Moringa is
caused by the destabilisation of negatively charged
colloids by cationic polyelectrolytes. Figure 5 shows
the residual turbidity and the zeta potential of the
kaolin suspension as a function of the dosage of a 5%
solution from non-shelled Moringa seeds.
It can be seen from this figure that the optimal
dosage of the natural coagulant corresponds to the
zeta potential of zero. At a dosage of 10ml/l
(500 mg/1) of this coagulant (non-shelled seeds), the
turbidity is reduced to 10 NTU. An overdosage of the
coagulant tends to reverse the zeta potential of the
kaolin suspension at around 4 m V . However, it
does not result in the restabilisation of the colloids,
as the residual turbidity remains low. The most likely
mechanisms involved in this coagulation activity are
adsorption and neutralisation of charges, or adsorption and bridging of destabilised particles (Weber,
1972; AWWA, 1991; Bratby, 1980; Attia, 1987). It is
difficult to identify which of these two mechanisms
apply, since they could, in fact, be taking place

aL

simultaneously (Bratby, 1980). As the ion-exchange

columns showed that positive charges were a prerequisite for coagulation to be initiated, and as the zeta
potential measurements showed that the optimal
dosage corresponded to zero zeta potential, the predominant mechanism of the coagulation with
Moringa appears to be adsorption and charge neutralisation.
In Fig. 6, where both shelled and non-shelled
Moringa seeds were used to treat a synthetic water of
an initial turbidity of 105 NTU, it can be noted [Fig.
6(a)] that the optimal dosage is 1 ml/l (50 mg/l) for
shelled seeds, while it is 10 times greater for nonshelled ones. This is to be expected since the active
proteins are less concentrated in the non-shelled seeds
extracts. It can also be seen for shelled Moringa seeds
that the residual turbidity increases when the coagulant dosage is increased beyond the optimal dosage.
This may be due to re-stabilisation cause by reversal
of colloidal charge due to excess of adsorption
(Bratby, 1980). This restabilization can be explained
by the protein in Moringa solution which, as reported
in Table 3, is 0.9% in case of shelled Moringa seeds
and only 0.3% in case of non-shelled ones.

Comparison of efficiency between Moringa and alum

Figure 6 shows the results of comparative coagulation tests between raw aqueous extracts of shelled
and non-shelled Moringa seeds, and also aluminum
sulphate.
As shown in Fig. 6(a), at coagulants dosages up to
10 ml/l residual turbidities in the case of alum and
Moringa are almost the same. It may be recalled that
with an initial turbidity of 426 NTU the shelled and
the non-shelled Moringa were equally effective as
coagulants as seen in Fig. 1, but with an initial
turbidity of 105 NTU, the dosage required for equal
turbidity removal is 10 times higher for the nonshelled Moringa than for the shelled ones. This is to
be expected since the protein concentration in the
non-shelled Moringa extract is less than half of the
other extract.
From Fig. 6(b) it can be seen that the conductivity
of the treated water increases to about 650ps/cm
using alum. The shelled and non-shelled Moringa
seeds do not affect the conductivity at all, which
remains more or less at its initial value of 150 # s/cm.
Such ionic strength using alum often causes corrosion
in water supply and distribution networks.
Figure 6(c) shows the variation of pH as a function
of coagulant dosage. Alum causes the pH to decrease
rapidly to around 4.2, whereas the two preparations
of Moringa seeds do not significantly alter the pH.
The low pH resulting from the use of alum can only
be compensated by addition of alkali like lime or
sodium hydroxide, unless the water has already sufficient alkalinity. By comparison, Moringa as a coagulant requires no pH adjustment.
Lastly, in Fig. 6(d) are plotted values of sludge
volumes produced as a function of coagulant dosage.
It can be seen that for a dosage of 10ml/l of
respective coagulant, the ratio of the volume of

Coagulation of turbid waters

120

~ Alum
NSMS
o SMS

100 ~,
[-.
Z

1000

(a)

--

709

(b)
* Alum
NSMS
o SMS

800 -

80
600 -60

"~..
o
t...)

l0

20

400

2OO

30

--

10

(c)

7 ~

-\

20

30

Dosage (ml/l)

Dosage (ml/1)
8

I
10

~, Alum
NSMS
o SMS

(d)
~+l+k
+- Alum
,
~ ~
NSMS
SMS

Q
4

(/3

4 --

3
0

I
10

I
20

]
30

Dosage (mill)

I
10

20

30

Dosage (ml/l)

Fig. 6. Comparison of coagulation efficiency of shelled (SMS), non-shelled (NSMS) Moringa seeds and
alum: variation of some water quality parameters vs dosage. (a) Turbidity, (b) Conductivity, (c) pH, and
(d) Sludge volume
sludge produced in the case of alum, shelled and
non-shelled Moringa seeds extract is respectively
5:2.5:1. The problem of sludge after Moringa-based
coagulation only depends on the nature of coagulated
materials. An additional advantage in this case is that
all Moringa by-products are organic and biodegradable.
CONCLUSION
The following conclusions can be drawn from this
on going study:

Moringa oleifera is an effective natural coagulant which can be used in water treatment in two
principal crude forms: shelled or non-shelled dry
seeds.
The action of Moringa oleifera as a coagulant lies
in the presence of water soluble cationic proteins in
the seeds.
These proteins are densely charged cationic dimers
with a molecular weight of about 13 kDa. Adsorption
and neutralization of charges are the main mechanisms of coagulation.

It is possible to extract an edible vegetable oil from

the Moringa seeds as well as the coagulant.
In coagulation, Moringa hardly affects the pH and
conductivity.
The volume of sludge produced is considerably less
in case of Moringa than in case of alum.
The only hurdle to the adoption of Moringa for
water and wastewater treatment seems to be the
adequate supply of the seeds. A solution to this
problem will be perhaps the intensive cultivation of
Moringa tree in tropical countries, exactly like coffee
or tea, two important cash crops grown only in
tropical regions but consumed all over the world.
Gene cloning can also be a possible alternative but
it can be very expensive.

Acknowledgement--The authors gratefully acknowledge the

financial support of the Programme Canadien des Bourses
de la Francophonie.

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