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



Copyright © 1995 Elsevier Science Ltd
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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

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
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.

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,
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

ion-exchange. or simply in distilled water. a model turbid water was prepared by adding kaolin to tap water. Attia. 42 paper and then through a 0. namely. 1988. of Moringa solutions were also tried for their coagulating activity. a d s o r p t i o n a n d bridging. 1962. The required amount of salt was added directly to the Moringa solutions. 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. Edzwald et al. Canada. Five grams of laboratory grade kaolin (Anachemia. The suspension was then filtered firstly through Whatman No. Characterization was done by conducting elemental analysis of the seeds. chloroform and hexane. Preparation of Moringa In this study. to the solvent under investigation.18H20 ] used in this study was of laboratory grade (Anachemia AC-405). the 20 min at 40 rpm for slow mixing and the 30 min of sedimentation. a cation-exchange resin. that is the content of carbon.000 Da. namely carboxymethyl cellulose (CM cellulose C-50). 1989. The maximum particle size remaining in the kaolin suspension was estimated to be about 2/~m by Stokes law. we refer to the four basic m e c h a n i s m s of classic coagulation theory. water was retained as solvent and the standard concentration of 5% by weight (crude powder/solvent) was adopted. Successive extractions were also conducted to study the compatibility of the exploitation of vegetable oil and the active agents in the coagulation process. ACS 093). compression of the double layer.05 M Tris-HCl buffer (pH of 7. where they were stored until used. 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. but is a stable suspension used here to study the mechanism of coagulation. Kawamura. The kaolin suspension was diluted using tap water to obtain any desired turbidity. since higher concentrations were too hard to filter.000 and 30. AC-5302) were added to a litre of water. namely. It may be added here that there is no standard method for conducting the jar test. 1972. A chronometer was then started to control the 2min at 100rpm for rapid mixing. "'Synthetic" turbid water For the purpose of our experiments. and an anion-exchange resin. 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.05 M phosphate buffer (pH of 7. acetone. nitrogen.45 t~m nylon membrane.ANSELMENDABIGENGESEREet a/. 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. Scopes. the green and dry pods. 10. 1987) and therefore will not be elaborated here. chemical precipitation and electrophoresis. Dialysis tubes (Spectra/Por) with molecular weight cutoff from 6000 to 8000Da. Dentel a n d Gosset. ultrafiltration. 1991b). 1981. in the appropriate proportion. The aluminum sulphate [AI2(SO4)3.. the shelled and non-shelled seeds. The protein content. This was followed by 30 min of sedimentation. 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. YMI0 and YM30. Coagulation activity of each fraction was tested as already described by the jar test. Only water was able to extract the active agents in the coagulation with Moringa. Coagulation activity of each Moringa extract was verified by the jar test. a n d then e n m e s h m e n t in the precipitate (Weber. Also determined were the protein.5). lyophilisation. Chemical analysis on Moringa Various tests were performed on Moringa oleifera in its various forms. namely diethylaminoethyl (DEAE Sephadex A-50) were employed. and the whole mixture was stirred for 30 min. 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). Comparative coagulation tests were run under the same conditions as described above but using 5% alum solution instead of Moringa..000Da (Spectrum Medical Industries) were used. Different solvents were tested for their capacity to extract the active ingredients. a d s o r p t i o n a n d neutralization o f charges. petroleum ether. using standard methods. 1987).0). resulting in an initial turbidity of 105 NTU. and from 12. . This model water does not represent real water in any country. and also the bark enveloping the seeds were tested for their coagulation potential. water.45 #m) and then redissolved in distilled water or in 0. These techniques are adequately described in numerous references (Jirgensons. the precipitate was separated by filtration on a membrane filter (0. which are permeable to compounds having respectively molecular weights less than 1000. The supernatant was carefully removed and stored in a plastic bottle. fat and sugar contents. the powder was added. In order to explain the m e c h a n i s m of coagulation with Moringa oleifera. Quebec. 1988. hydrogen and oxygen. Following homogenization for 30 min using a magnetic stirrer. MATERIALS AND METHODS Coagulants used The Moringa oleifera used as a coagulant in this study comes from Burundi in Central Africa. Solids obtained by lyophilisation and by evaporation at 20°C and also at 105°C. In order to determine the nature of the charges carried by the coagulant active proteins. Precipitation of active proteins from Moringa solutions was carried out using 80-100% saturated solutions of ammonium sulphate (Anachemia.4) or in 0. For example. Benefield et al. 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. Lyophilisation of various fractions of Moringa solutions was carried out using a Labconco freeze dryer (model 4. After a first series of tests. The suspension was stirred for 30 min and then allowed to settle for 24hs.000 to 14.05 M phosphate buffer (pH of 7. Coagulation tests were conducted using the jar test equipment of Phipps & Bird having a base floc illuminator.0). 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. Ultrafiltration was carried out using a Millipore cell (GS Amicon) and the membranes YM2. One litre of the synthetic water was used for the test in each of the 6 beakers placed on the base illuminator. 1982. Purification and characterization of active agents Active agents were purified and characterized using different techniques. The extraction of the active ingredients was carried out as follows: the raw material was ground to powder in a domestic blender. namely: dialysis. the principal objective being the determination of its general composition and its potential use as a natural coagulant in water and wastewater treatment. Franks. 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. Coagulation test The jar test has been and is still the method widely used to evaluate coagulation-flocculation processes (Hudson.

for highly turbid waters. if the turbidity removal was below 30% the result was described as "absent". Parameters analyzed The following parameters were analyzed in the raw and treated water using Standard Methods (APHA. we found that the latter was almost as good as the former as a coagulant. During the experiment.Coagulation of turbid waters Table I. Turbidity measurements were conducted using a Hellige turbidimeter (series 12520). Sludge volume was measured using Imhoff cones. lyophilised and then stored at ambient temperature. Coagulating activity of shelled (SMS) and nonshelled (NSMS) Moringa seeds . If the dry seeds are removed and ground separately. This means that when treating highly turbid waters. 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. 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. Considerable turbidity (80-90%) was removed after sedimentation in the jar test. Conductivity was measured also with the same brand name instrument. Zeta potential was measure with a zetameter Zetasizer II of Malvern. On the contrary. This is a qualitative assessment of the coagulating capacity of Moringa pods and seeds. We also studied the effect of the concentration of Moringa solution on the coagulation activity.0). From Table 1. sludge volume and zeta potential. it is also to be noted that the bark around the seed has no coagulation properties. it is not necessary to separate the shell from the seed. Comparing the shelled and non-shelled Moringa seeds. the pods should be allowed to dry completely on the 705 q ~" 400 [-Z 300 * Shelled Moringa seeds (SMS) O Non-shelled Moringa seeds (NSMS) 200 I00 10 20 30 Dosage (mill) Fig. More concentrated Moringa solutions are better because small volumes are required to get the optimal dosage. The extraction in water of the whole dry pods did not give any positive result. all preparations based on green pods of Moringa do not possess any coagulation activity. it is advantageous to use the nonshelled seeds in order to save the labour and energy required to separate one from the other. 1992): pH value. 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. Removing shells from Moringa seeds is a tedious process consuming time and expense. whereas the residual solids do not show any sign of coagulation activity. 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. 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. with 5% solutions of shelled and non-shelled Moringa seeds.05 M phosphate buffer (pH of 7. the filtered and unfiltered solution of Moringa seeds has coagulant properties. with a concentration gradient 0-1 M NaCI in the same buffers. turbidity. shelled seeds were more effective coagulant then non-shelled ones. On an industrial scale. for an initial turbidity of 426 N T U . Therefore. for Moringa to serve as a coagulant one cannot directly pulverize the dry pods. Coagulation activity is present in shelled and in non-shelled seeds but only after the pods are dry. The pH value was measured with a Hach-one pH meter. The conclusion from the foregoing results is that the active agents in the coagulation with Moringa are water soluble materials. Studies on the purified proteins were carried out using an S-18 reverse phase column on a Beckman HPLC system. RESULTS AND DISCUSSION plant. 1. as will be discussed later. Purified active extracts were desalted against water. Figure 2 shows turbidity removal after coagulation with different concentrations of non-shelled Moringa seeds solutions. This means that for exploitation of Moringa seeds as a coagulant. In this case the coagulation activity was qualified as "present". Figure 1 shows the results of coagulation of a model water having an initial turbidity of 426 N T U . When the initial turbidity was lowered to 1 0 5 N T U however. Similarly. the coagulation activity was found in the seeds. if necessary. coagulant property was manifested by formation of visible flocs. As can be seen from the Table 1. conductivity. before using Moringa seeds as a coagulant. easily recognized by naked eye following flow agitation of the turbid water.

6 24.9 27. As can be seen from Table 2. Results of elemental analysis of shelled and nonshelled Moringa seeds are presented in Table 2. and 21% in the non-shelled seeds. 1974) use the proportionality factor of 6. 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. Organic matter consists of the six principal elements. the nitrogen content of the whole non-shelled Moringa seeds can be estimated from Table 2 as follows: (100"6.0 -1. hexane. In our studies.0 26. hexane and chloroform are mainly vegetable oils. l 26.3 Non-sheUed --Powder --Solution --Residual solids 27.3 absent absent absent absent present absent .3 34.8%. along with their respective coagulating activity is shown in Table 4.0 53.ANSELME NDABIGENGESERE el al.4 27.6 0.3 5.7 0. Using the hypothesis that the Proteins (%) Lipids (%) Carbohydrates (%) Shelled --Powder --Solution --Residual solids 36. carbon tetrachloride. This is why the coagulant dosage is expressed mostly in ml/1 throughout this study. lipids and sugars in Moringa seeds - i~. acetone. The principal groups of the above constituents are proteins. phosphorus (P) and sulphur (S). Moringa seeds preparation * 0.5% 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. oxygen (O).1 + 70"5)/(2"100) = 4. Carbohydrates (as oligosaccharides) represent 5% of the shelled seeds whereas they are 5.1 0. namely carbon (C).4 25. Nitrogen is a common component in all proteins and free amino acids (Rogers. The remaining 31% consists of oxygen and trace elements. 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. hydrogen (H).8 50. Coagulating activity of non-shelled Moringaseeds at different concentration in aqueous solution all the four concentrations studied. we found that the shelled Moringa seeds contain 55% carbon. The extracts with petroleum ether.3% in case of shelled seeds and to 27. 706 120 - Table 3. Some authors (Strong. water and methanol. 1991).25 to estimate the protein content of organic matter from the available values of nitrogen. lipids and carbohydrates. 27% in the non-shelled ones.5% "N 0 I I I 50 100 150 Dosage (mill) Fig.4 2 I. carbohydrates. The shelled Moringa seeds contain about 37% proteins.5 --- nitrogen is contained only in the cotyledon.9 29. 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. whereas there is close to 35% of lipids in the shelled Moringa seeds. General composition of Moringa oleifera Data presented in this section are simple averages of at least two analyses.5 5. the non-shelled Moringa seeds trail closely the shelled Moringa seeds in all the elements analyzed. 1 0.3% for non-shelled ones.8 8. Elemental analysis of Moringa (% by mass) Sample Shelled Moringa seeds (SMS) Non-Shelled Moringa seeds (NSMS) N (%) C (%) H (%) 6. 8.3 26. The 5% Moringa solution was retained as the standard in this work. Only the water extract possess coagulation activity.3 7. The lipids are not soluble in water. 2. Table 4. lignins and free amino acids (Jirgensons. It is clearly possible to extract oil first and then use the aqueous extract as a coagulant. which is very close to the measured nitrogen content of 5% for nonshelled seeds.1 54. 1962). all of them being inferior by a small margin however. The percentage of extractable matter of Moringa seeds using various solvents like ether. lipids.3 5. Coagulation activity of extracts from Moringa with different solvents Solvent Table 2. nitrogen (N).5% hydrogen and 6% nitrogen. 1962). It appears that almost all the proteins and amino acids are in the seed cotyledons. Percentage (by mass) of proteins. Table 3 shows the results of the analysis of Moringa oleifera into proteins. whereas the extract with acetone and methanol are mainly carbohydrates (Jirgensons.7 Petroleum ether Hexane Chloroform Acetone Water Methanol Quantity of extractable matters (% by mass) Coagulation activity 33. this is why their content in the residual solids increases to 50.

In reducing conditions. Gassen et al. and he separated two active fractions (MO1 and MO2) on columns. Elution of Moringa proteins from a CM cellulose column.000 Da. uttrafiltration tests showed that the active ingredients were retained on the 10 kDa membranes.05 0. 1991). The photograph shows the migration of a series of molecular weight standards. Precipitation tests with ammonium sulphate showed that the precipitate has excellent coagulant properties. and the Moringa proteins. even after 6 months of storage without any special precaution as to the pH and temperature. 1990.6 ¢9 t~ e- 0. The Moringa proteins are stable in this dried form and remain so.6-1.4.5 kDa. 3. Experiments using polyacrylamide gel isoelectric focusing in a pH gradient from 3 to 11. However. 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.P A G E tests. 4. ranging from 14 to 2000 kDa.2 0 0 50 100 150 200 Z 0 Elution volume (ml) Fig. 3.. This means that isolation and purification are relatively easy 0.8 o. lipids and carbohydrates. A2 and A3) which is consistent with their being at least two different monomers which can associate at random.5 kDa under reducing and of 13 kDa under non-reducing conditions. with the CM cellulose.. Similarly. purified by CM-cellulose to the gel. Dialysis experiments showed that coagulation activity stayed inside of the dialysis tubes with molecular weight cutoff from 12. but passed through the 30 kDa membranes.Coagulation of turbid waters MW kDa 9 7 ~ 4 6 ~ 3 1 ~ 2 1 ~ 14-- Std B C Fig. A2 and A3 represent discrete factions with coagulating properties.000. as shown in Fig. Std: molecular weight standards Characterization of the active agents in coagulation with Moringa The aqueous extract of Moringa is far from being pure. Gassenschmidt et al. AI. 10/ag of polled coagulating proteins. 1987). Lyophilisation of crude Moringa solution and also the purified Moringa proteins results in a white powder. in presence and absence of mercaptoethanol. (C) under non-reducing conditions.000 to 14. When 5% Moringa solution was applied to DEAE cellulose at pH of 7.. 4. Later. as shown in Fig.20 -- -'] 1.. which is soluble in water and its solution has very good coagulant properties. the active proteins did not bind. W R 29'2 U 707 Determination of the molecular weight of the proteins extracted from Moringa seeds by molecular sieving method showed molecular weight of 6. Ion-exchange column experiments showed that the active proteins carry positive charges. . Ammonium sulphate precipitates are known to be proteins (Scopes. It is a solution consisting principally of proteins. SDS PAGE separation on Moringa proteins. our results confirm the suggestion by Gassen et al.4 0. 1984.. (B) under reducing conditions (1 mM mercaptoethanol). the active proteins were bound and could be eluted in a very pure state at 0. thus clearly demonstrating their highly cationic nature.5 kDa. Together these results showed that the native protein was a dimeric of 13 kDa with subunits of about 6. using the purified Moringa proteins from the CM cellulose column showed that these proteins had isoelectric points between 10 and 11. Gassenschmidt (1991) suggested the sequence of the two polypeptides. There were three active peptides isolated from this column (A1. Gassen (1990) purified the two fractions and found that they are proteins of 6500Da. An identical result was obtained by S D S . A series of experiments were carried out to characterize the coagulating and flocculating agents of this mixture. Both monomers and dimers retain their coagulant properties.15 . GradientNaCl // " //1 o 1. (1990) that the active proteins have a molecular weight of 6.2 / DO 280 nm 0. The supernatant did not possess any coagulation activity. The subunits are liked by S-S bonds.lo c 0.0 ~ 0. This result supports the previous works (Fink.0 M NaC1.

t ~ k . fast spin evaporation may be the most economically feasible approach. 6(b) it can be seen that the conductivity of the treated water increases to about 650ps/cm using alum. and as the zeta potential measurements showed that the optimal dosage corresponded to zero zeta potential. At a dosage of 10ml/l (500 mg/1) of this coagulant (non-shelled seeds). However. Hence coagulation of the kaolin suspension using Moringa is caused by the destabilisation of negatively charged colloids by cationic polyelectrolytes. This is to be expected since the active proteins are less concentrated in the non-shelled seeds extracts. Figure 6(c) shows the variation of pH as a function of coagulant dosage. . unless the water has already sufficient alkalinity. 6(a)] that the optimal dosage is 1 ml/l (50 mg/l) for shelled seeds. 1991. be taking place aL simultaneously (Bratby. the ratio of the volume of . where both shelled and non-shelled Moringa seeds were used to treat a synthetic water of an initial turbidity of 105 NTU.4 6 inV. From Fig.-to -20 = -30 2 r~ 40 \ 0 0 _ I I I 10 20 30 -40 -50 Dosage (ml/l) Fig. This restabilization can be explained by the protein in Moringa solution which. . as reported in Table 3. It can be seen that for a dosage of 10ml/l of respective coagulant. 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. it can be noted [Fig. As the ion-exchange columns showed that positive charges were a prerequisite for coagulation to be initiated. 1980). the kaolin particles were charged negatively. z A--"~" . . it does not result in the restabilisation of the colloids. 1988). Attia. Bratby. ~ 0 . . An overdosage of the coagulant tends to reverse the zeta potential of the kaolin suspension at around ÷ 4 m V . 1987). Hence. while it is 10 times greater for nonshelled ones. 1. 6(d) are plotted values of sludge volumes produced as a function of coagulant dosage.ANSELMENDABIGENGESERE et 708 120 10 . 1980).9% in case of shelled Moringa seeds and only 0./ [-. Lastly. This means that at a pH of around 7. as the residual turbidity remains low. 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. the active agents in coagulation with Moringa are water soluble cationic proteins. By comparison. 5. is 0.2. 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. confirming that even if the solution is a heterogenous complex mixture. The zeta potential of a 5% solution of non-shelled Moringa seeds was found to be + 6 mV. The shelled and non-shelled Moringa seeds do not affect the conductivity at all. at coagulants dosages up to 10 ml/l residual turbidities in the case of alum and Moringa are almost the same. 1972. This is to be expected since the protein concentration in the non-shelled Moringa extract is less than half of the other extract. 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. This may be due to re-stabilisation cause by reversal of colloidal charge due to excess of adsorption (Bratby. in fact. in Fig. It can be seen from this figure that the optimal dosage of the natural coagulant corresponds to the zeta potential of zero. In Fig. 1980. 6(a). the predominant mechanism of the coagulation with Moringa appears to be adsorption and charge neutralisation. The most likely mechanisms involved in this coagulation activity are adsorption and neutralisation of charges. which remains more or less at its initial value of 150 # s/cm. whereas the two preparations of Moringa seeds do not significantly alter the pH. since they could. slow evaporation of Moringa solution at ambient temperature and also at about 105°C resulted in some denaturation of the proteins which became difficult to solubilize and less efficient in coagulation. Alum causes the pH to decrease rapidly to around 4. for best results in the preparation of Moringa as a coagulant. and also aluminum sulphate. However. The low pH resulting from the use of alum can only be compensated by addition of alkali like lime or sodium hydroxide. The zeta potential of the synthetic water was . AWWA. but with an initial turbidity of 105 NTU. 6. t -t 80 ~* AAI' • Zeta i~ * NTU ~3 . the dosage required for equal turbidity removal is 10 times higher for the nonshelled Moringa than for the shelled ones. if not freeze drying. or adsorption and bridging of destabilised particles (Weber. Moringa as a coagulant requires no pH adjustment. Mechanism of coagulation with Moringa As mentioned before. the positive charges are predominant in the solution. It is difficult to identify which of these two mechanisms apply. the turbidity is reduced to 10 NTU.3% in case of non-shelled ones. It can also be seen for shelled Moringa seeds that the residual turbidity increases when the coagulant dosage is increased beyond the optimal dosage. Such ionic strength using alum often causes corrosion in water supply and distribution networks. As shown in Fig.

The volume of sludge produced is considerably less in case of Moringa than in case of alum. ~ ~ NSMS SMS 6 Q 4 (/3 2 4 -- O 3 0 I 10 I 20 ] 30 Dosage (mill) I 10 20 30 Dosage (ml/l) Fig. Magara Y.. Comparison of coagulation efficiency of shelled (SMS). [-. The action of Moringa oleifera as a coagulant lies in the presence of water soluble cationic proteins in the seeds. Acknowledgement--The authors gratefully acknowledge the financial support of the Programme Canadien des Bourses de la Francophonie. IWSA/IA WPRC Joint . The problem of sludge after Moringa-based coagulation only depends on the nature of coagulated materials. A solution to this problem will be perhaps the intensive cultivation of Moringa tree in tropical countries.) 0 l0 20 400 - / 2OO 30 -- 6 5 10 (c) 7 ~ -\ ~ • ~ I I 20 30 Dosage (ml/l) Dosage (ml/1) 8 I 10 • 8 ~.Coagulation of turbid waters 120 - ~ Alum • NSMS o SMS 100 ~. Z 1000 (a) -- 709 (b) * Alum • NSMS o SMS 800 - * 80 600 -60 "~. The only hurdle to the adoption of Moringa for water and wastewater treatment seems to be the adequate supply of the seeds. An additional advantage in this case is that all Moringa by-products are organic and biodegradable. In Proc. It is possible to extract an edible vegetable oil from the Moringa seeds as well as the coagulant. Moringa hardly affects the pH and conductivity.. two important cash crops grown only in tropical regions but consumed all over the world. (c) pH. Adsorption and neutralization of charges are the main mechanisms of coagulation.. REFERENCES Aizawa T.Alum . o t. (a) Turbidity. 6. non-shelled (NSMS) Moringa seeds and alum: variation of some water quality parameters vs dosage. and Musashi (1990) Problems with introducing synthetic polyelectrolytes coagulants into the water purification. These proteins are densely charged cationic dimers with a molecular weight of about 13 kDa. In coagulation.5:1.. shelled and non-shelled Moringa seeds extract is respectively 5:2. and (d) Sludge volume sludge produced in the case of alum. Alum • NSMS o SMS ~ (d) ~+l+k +. (b) Conductivity. 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. Gene cloning can also be a possible alternative but it can be very expensive. exactly like coffee or tea.

D. 82. and Weand B. K.J.). Chem. P. Miller R. (1988) Characterisation o f Proteins. J. Wiley. 75. E.. H. (1986) Proper use of African Natural Coagulants for Rural Water Supplies: Research in the Sudan and a Guide for New Projects. Oxford. Strong F. Washington. and Koch G. Universit6 du Burundi. Wat.. Univ. L. Flocculation.. Wat. and Selleck R. Edzwald J. Bujumbura. (1991). Am. Wagner E.. Eschborn. Hudson H. Chemical properties of flocculating active proteins from Moringa oleifera. D. (1980) Wirkstoffe zur trinkwasseraufbereitung aus samen von moringa oleifera. P. Dentel S. In Compte Rendu du s$minaire sur les Mbthodes appropri~es de traitement de I' eau dans le milieu rural (Edited by Kaiser F. R. Bujumbura. 244-247. Sedimentation and Floatation. War. Universit6 du Burundi. Wks Assoc. J. Calif. (1985) Toxicity and mutagenicity evaluation of water coagulated with Moringa oleifera seed preparations using fish. J. Germany.710 ANSELMENDABIGENGESEREet al. Methods in Plant Biochemistry. 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